THERAPEUTIC AND DIAGNOSTIC METHODS AND COMPOSITIONS BASED ON NOTCH PROTEINS AND NUCLEIC ACIDS

Abstract: The present invention relates to therapeutic and diagnostic methods and compositions based on Notch proteins and nucleic acids. Figure 17 displays the sequences of human Notch DNA and the encoded human Notch protein. The invention provides for treatment of disorders of cell fate or differentiation by administration of a therapeutic compound of the invention. Such therapeutic compounds (termed herein "Therapeutics") include Notch proteins and analogs and derivatives (including fragments) thereof, antibodies thereto, nucleic acids encoding the Notch proteins, analogs, or derivatives, Notch antisense nucleic acids, as well as toporythmic proteins and derivatives which bind to or otherwise interact with Notch proteins, their encoding nucleic acids or antibodies. In a preferred embodiment, a Therapeutic of the invention is administered to treat a cancerous condition, or to prevent progression from a pre-neoplastic or non-malignant state into a neoplastic or a malignant state.

Description: THERAPEUTIC AND DIAGNOSTIC METHODS AND COMPOSITIONS BASED ON NOTCH PROTEINS AND NUCLEIC ACIDS This application is a continuation-in-part of copending application "V - Serial No. 08/083,590 filed June 25, 1993, which is a continuation-in-part of both application Serial No. 07/955,012 filed September 30, 1992, now abandoned, and copending application Serial No. 07/879,038 filed April 30, 1992, each of which is incorporated by reference herein in its entirety. This invention was made in part with government support under JQ grant numbers GM 29093 and NS 26084 awarded by the National Institutes of Health. The government has certain rights in the invention. 1. INTRODUCTION The present invention relates to therapeutic compositions J5 comprising Notch proteins, analogs and derivatives thereof, antibodies thereto, nucleic acids encoding the Notch proteins, derivatives or analogs, Notch antisense nucleic acids, and toporythmic proteins which bind to Notch and their nucleic acids and antibodies. Therapeutic and diagnostic methods are also provided. 20 2. BACKGROUND OF THE INVENTION 2.1. THE NOTCH GENE AND PROTEIN Null mutations in any one of the zygotic neurogenic loci ~ Notch (N), Delta (DI), mastermind (mam). Enhancer of Split (E(spl). neuralized (neu). 2 and big brain (bib) —result in hypertrophy of the nervous system at the expense of ventral and lateral epidermal structures. This effect is due to the misrouting of epidermal precursor cells into a neuronal pathway, and implies that neurogenic gene function is necessary to divert cells within the neurogenic region from a neuronal fate to an epithelial fate. Studies that assessed the effects of laser 30 ablation of specific embryonic neuroblasts in grasshoppers (Doe and Goodman 1985, Dev. Biol. I ll, 206-219) have shown that cellular interactions between neuroblasts and the surrounding accessory cells serve to inhibit these accessory 35 cells from adopting a neuroblast fate. Together, these genetic and developmental observations have led to the hypothesis that the protein products of the neurogenic loci function as components of a cellular interaction mechanism necessary for proper epidermal development (Artavanis-Tsakonas, 1988, Trends Genet. 4, 95- 100). Sequence analyses (Wharton et al., 1985, Cell 43, 567-581 ; Kidd et al., 1986, Mol. Cell. Biol. 6, 3094-3108; Vassin et al., 1987, EMBO J. 6, 3431-3440; Kopczynski et al., 1988, Genes Dev. 2, 1723-1735) have shown that two of the neurogenic loci, Notch and Delta, appear to encode transmembrane proteins that span the membrane a single time. The Drosophila Notch gene encodes a — 300 kd protein (we use "Notch" to denote this protein) with a large N-terminal extracellular domain that includes 36 epidermal growth factor (EGF)- like tandem repeats followed by three other cysteine-rich repeats, designated Notch/lin-12 repeats (Wharton et al., 1985, Cell 43, 567-581 ; Kidd et al., 1986, Mol. Cell Biol. 6, 3094-3108; Yochem et al., 1988, Nature 335, 547-550). The sequences of Xenopus (Coffman et al., 1990, Science 249: 1438-1441) and a human Notch homolog termed TAN-1 (Ellisen et al., 1991 , Cell 66:649-661) have also been reported. Delta encodes a ~ 100 kd protein (we use "Delta" to denote DLZM, the protein product of the predominant zygotic and maternal transcripts; Kopczynski et al. , 1988, Genes Dev. 2, 1723-1735) that has nine EGF-like repeats within its extracellular domain (Vassin et al., 1987, EMBO J. 6, 3431- 3440; Kopczynski et al., 1988, Genes Dev. 2, 1723-1735). Although little is known about the functional significance of these repeats, the EGF-like motif has been found in a variety of proteins, including those involved in the blood clotting cascade (Furie and Furie, 1988, Cell 53, 505-518). In particular, this motif has been found in extracellular proteins such as the blood clotting factors IX and X (Rees et al., 1988, EMBO J. 7, 2053-2061; Furie and Furie, 1988, Cell 53, 505- 518), in other Drosophila genes (Knust et al., 1987, EMBO J. 761-766; Rothberg et al., 1988, Cell 55, 1047-1059), and in some cell-surface receptor proteins, such as thrombomodulin (Suzuki et al., 1987, EMBO J. 6, 1891-1897) and LDL receptor (Sudhof et al., 1985, Science 228, 815-822). A protein binding site has been mapped to the EGF repeat domain in thrombomodulin and urokinase (Kurosawa et al., 1988, J. Biol. Chem 263, 5993-5996; Appella et al., 1987, J. Biol. Chem. 262, 4437-4440). An intriguing array of interactions between Notch and Delta mutations has been described (Vassin, et al., 1985, J. Neurogenet. 2, 291-308; Shepard et al., 1989, Genetics 122, 429-438; Xu et al., 1990, Genes Dev., 4, 464-475). A number of genetic studies (summarized in Alton et al., 1989, Dev. Genet. 10, 261-272) has indicated that the gene dosages of Notch and Delta in relation to one another are crucial for normal development. A 50% reduction in the dose of Delta in a wild-type Notch background causes a broadening of the wing veins creating a "delta" at the base (Lindsley and Grell, 1968, Publication Number 627, Washington, D.C, Carnegie Institute of Washington). A similar phenotype is caused by a 50% increase in the dose of Notch in a wild-type Delta background (a "Confluens" phenotype; Welshons, 1965, Science 150, 1122- 1129). This Delta phenotype is partially suppressed by a reduction in the Notch dosage. Work has shown that lethal interactions between alleles that correlate with alterations in the EGF-like repeats in Notch can be rescued by reducing the dose of Delta (Xu et al., 1990, Genes Dev. 4, 464-475). Xu et al. (1990, Genes Dev. 4, 464-475) found that null mutations at either Delta or mam suppress lethal interactions between heterozygous combinations of certain Notch alleles, known as the Abruptex (Ax) mutations. Ax alleles are associated with missense mutations within the EGF-like repeats of the Notch extracellular domain (Kelley et al., 1987, Cell 51, 539-548; Hartley et al., 1987, EMBO J. 6, 3407-3417). Recent studies have shown that Notch and Delta, and Notch and Serrate, directly interact on the molecular level (Fehon et al., 1990, Cell 61:523- 534; Rebay et al., 1991, Cell 67:687-699). Notch is expressed on axonal processes during the outgrowth of embryonic neurons (Johansen et al. , 1989, J. Cell Biol. 109:2427-2440; Kidd et al., 1989, Genes Dev. 3:1113-1129; Fehon et al. , 1991 , J. Cell Biol. 113:657-669). A study has shown that certain Ax alleles of Notch can severely alter axon pathfinding during sensory neural outgrowth in the imaginal discs, although it is not yet known whether aberrant Notch expression in the axon itself or the epithelium along which it grows is responsible for this defect (Palka et al., 1990, Development 109, 167-175). 2.2. CANCER A neoplasm, or tumor, is a neoplastic mass resulting from abnormal uncontrolled cell growth, which may cause swelling on the body surface, and which can be benign or malignant. Benign tumors generally remain localized. Malignant tumors are collectively termed cancers. The term "malignant" generally means that the tumor can invade and destroy neighboring body structures and spread to distant sites to cause death (for review, see Robbins and Angell, 1976, Basic Pathology, 2d Ed., W.B. Saunders Co., Philadelphia, pp. 68-122). Effective treatment and prevention of cancer remains a long-felt need, and a major goal of biomedical research. 3. SUMMARY OF THE INVENTION The present invention relates to therapeutic and diagnostic methods and compositions based on Notch proteins and nucleic acids. The invention provides for treatment of disorders of cell fate or differentiation by administration of a therapeutic compound of the invention. Such therapeutic compounds (termed herein "Therapeutics") include: Notch proteins and analogs and derivatives (including fragments) thereof; antibodies thereto; nucleic acids encoding the Notch proteins, analogs, or derivatives; Notch antisense nucleic acids; as well as toporythmic proteins and derivatives which bind to or otherwise interact with Notch proteins, and their encoding nucleic acids and antibodies. In a preferred embodiment, a Therapeutic of the invention is administered to treat a cancerous condition, or to prevent progression from a pre-neoplastic or non-malignant state into a neoplastic or a malignant state. In other specific embodiments, a Therapeutic of the invention is administered to treat a nervous system disorder or to promote tissue regeneration and repair. In one embodiment, Therapeutics which antagonize, or inhibit, Notch function (hereinafter "Antagonist Therapeutics") are administered for therapeutic effect; disorders which can be thus treated can be identified by in vitro assays such as described in Section 5.1, infra. Such Antagonist Therapeutics include but are not limited to Notch antisense nucleic acids, anti-Notch neutralizing antibodies, and competitive inhibitors of Notch protein-protein interactions (e.g. , a protein comprising Notch ELR-11 and ELR-12 and derivatives thereof), all as detailed infra. In another embodiment, Therapeutics which promote Notch function (hereinafter "Agonist Therapeutics") are administered for therapeutic effect; disorders which can thus be treated can be identified by in vitro assays such as described in Section 5.1, infra. Such Agonist Therapeutics include but are not limited to Notch proteins and derivatives thereof comprising the intracellular domain, and proteins that interact with Notch (e.g. , a protein comprising a Delta sequence homologous to Drosophila Delta amino acids 1-230 (see Figure 1 and SEQ ID NO:2), or comprising a Serrate sequence homologous to Drosophila Serrate amino acids 79-282 (see Figure 5 and SEQ ID NO:4)). Disorders of cell fate, in particular hyperproliferative (e.g. , cancer) or hypoproliferative disorders, involving aberrant or undesirable levels of expression or activity of Notch protein can be diagnosed by detecting such levels, as described more fully infra. In a preferred aspect, a Therapeutic of the invention is a protein consisting of at least a fragment (termed herein "adhesive fragment") of the proteins encoded by toporythmic genes which mediates binding to Notch proteins or adhesive fragments thereof. Toporythmic genes, as used herein, shall mean the genes Notch. Delta, and Serrate, as well as other members of the Delta/Serrate family which may be identified by virtue of sequence homology or genetic interaction, and in general, members of the "Notch cascade" or the "Notch group" of genes, which are identified by molecular interactions (e.g. , binding in vitro) or genetic interactions (as detected phenotypically, e.g. , in Drosophila). In another aspect, the invention is directed to human Notch proteins; in particular, that encoded by the hN homolog, and proteins comprising the extracellular domain of the protein and subsequences thereof. Nucleic acids encoding the foregoing, and recombinant cells are also provided. 3.1. DEFINITIONS As used herein, the following terms shall have the meanings indicated: AA = amino acid EGF = epidermal growth factor ELR = EGF-like (homologous) repeat IC = intracellular PCR = polymerase chain reaction As used herein, underscoring the name of a gene shall indicate the gene, in contrast to its encoded protein product which is indicated by the name of the gene in the absence of any underscoring. For example, "Notch" shall mean the Notch gene, whereas "Notch" shall indicate the protein product of the Notch gene. 4. DESCRIPTION OF THE FIGURES Figure 1. Primary Nucleotide Sequence of the Delta cDNA Dll (SEQ ID NO:l) and Delta amino acid sequence (SEQ ID NO:2). The DNA sequence of the 5'-3' strand of the Dll cDNA is shown, which contains a number of corrections in comparison to that presented in Kopczynkski et al. (1988, Genes Dev. 2:1723-1735). Figure 2. Notch Expression Constructs and the Deletion Mapping of the Delta/Serrate Binding Domain. S2 cells in log phase growth were transiently transfected with the series of expression constructs shown; the drawings represent the predicted protein products of the various Notch deletion mutants created. All expression constructs were derived from construct #1 pMtNMg. Transiently transfected cells were mixed with Delta expressing cells from the stably transformed line L49-6-7 or with transiently transfected Serrate expressing cells, induced with CuS0 4 , incubated under aggregation conditions and then scored for their ability to aggregate using specific antisera and immunofluorescence microscopy. Aggregates were defined as clusters of four or more cells containing both Notch and Delta/Serrate expressing cells. The values given for % Aggregation refer to the percentage of all Notch expressing cells found in such clusters either with Delta (DI) (left column) or with Serrate (Ser) (right column). The various Notch deletion constructs are represented diagrammatically with splice lines indicating the ligation junctions. Each EGF repeat is denoted as a stippled rectangular box and numbers of the EGF repeats on either side of a ligation junction are noted. At the ligation junctions, partial EGF repeats produced by the various deletions are denoted by open boxes and closed brackets (for example see #23 ΔCla+EGF(10-12)). Constructs #3-13 represent the Clal deletion series. As diagrammed, four of the Clal sites, in repeats 7, 9, 17 and 26, break the repeat in the middle, immediately after the third cysteine (denoted by open box repeats; see Figure 3 for further clarification), while the fifth and most 3' site breaks neatly between EGF repeats 30 and 31 (denoted by closed box repeat 31; again see Figure 3). In construct #15 split, EGF repeat 14 which carries the split point mutation, is drawn as a striped box. In construct #33 ΔCla+XEGF(10-13), the Xenopus Notch derived EGF repeats are distinguished from Drosophila repeats by a different pattern of shading. SP, signal peptide; EGF, epidermal growth factor repeat; N, Notch/lin- 12 repeat; TM, transmembrane domain; cdclO, cdclO/ankyrin repeats; PA, putative nucleotide binding consensus sequence; opa, polyglutamine stretch termed opa; DI, Delta; Ser, Serrate. Figure 3. Detailed Structure of Notch Deletion Constructs #19-24: Both EGF Repeats 11 and 12 are Required for Notch-Delta Aggregation. EGF repeats 10-13 are diagrammed at the top showing the regular spacing of the six cysteine residues (C). PCR products generated for these constructs (names and numbers as given in Figure 2) are represented by the heavy black lines and the exact endpoints are noted relative to the various EGF repeats. Ability to aggregate with Delta is recorded as (+) or (-) for each construct. The PCR fragments either break the EGF repeats in the middle, just after the third cysteine in the same place as four out of the five Clal sites, or exactly in between two repeats in the same place as the most C-terminal Clal site. Figure 4. Comparison of Amino Acid Sequence of EGF Repeats 11 and 12 from Drosophila and Xenopus Notch. The amino acid sequence of EGF repeats 11 and 12 of Drosophila Notch (SEQ ID NO: 14) (Wharton et al. , 1985, Cell 43:567-581; Kidd et al., 1986, Mol. Cell Biol. 6:3094-3108) is aligned with that of the same two EGF repeats from Xenopus Notch (SEQ ID NO: 15) (Coffman et al., 1990, Science 249: 1438-1441). Identical amino acids are boxed. The six conserved cysteine residues of each EGF repeat and the Ca + + binding consensus residues (Rees et al. , 1988, EMBO J. 7:2053-2061) are marked with an asterisk (*). The leucine to proline change found in the Xenopus PCR clone that failed to aggregate is noted underneath. Figure 5. Nucleic Acid Sequence Homologies Between Serrate and Delta. A portion of the Drosophila Serrate nucleotide sequence (SEQ ID NO:3), with the encoded Serrate protein sequence (SEQ ID NO:4) written below (Fleming et al., 1990, Genes & Dev. 4:2188-2201 at 2193-94) is shown. The four regions showing high sequence homology with the Drosophila Delta sequence are numbered above the line and indicated by brackets. The total region of homology spans nucleotide numbers 627 through 1290 of the Serrate nucleotide sequence (numbering as in Figure 4 of Fleming et al. , 1990, Genes & Dev. 4:2188-2201). Figure 6. Schematic Diagram of Human Notch Clones. A schematic diagram of human Notch is shown. Heavy bold-face lines below the diagram show that portion of the Notch sequence contained in each of the four cDNA clones. The location of the primers used in PCR, and their orientation, are indicated by arrows. Figure 7. Human Notch Sequences Aligned with Drosophila Notch Sequence. Numbered vertical lines correspond to Drosophila Notch coordinates. Horizontal lines below each map show where clones lie relative to stretches of sequence (thick horizontal lines). Figure 8. Nucleotide Sequences of Human Notch Contained in Plasmid cDNA Clone hN2k. Figure 8A: The DNA sequence (SEQ ID NO:5) of a portion of the human Notch insert is shown, starting at the EcoRI site at the 3' end, and proceeding in the 3' to 5' direction. Figure 8B: The DNA sequence (SEQ ID NO:6) of a portion of the human Notch insert is shown, starting at the EcoRI site at the 5' end, and proceeding in the 5' to 3' direction. Figure 8C: The DNA sequence (SEQ ID NO:7) of a portion of the human Notch insert is shown, starting 3' of the sequence shown in Figure 8B, and proceeding in the 5' to 3' direction. The sequences shown are tentative, subject to confirmation by determination of overlapping sequences. Figure 9. Nucleotide Sequences of Human Notch Contained in Plasmid cDNA clone hN4k. Figure 9A: The DNA sequence (SEQ ID NO: 8) of a portion of the human Notch insert is shown, starting at the EcoRI site at the 5' end, and proceeding in the 5' to 3' direction. Figure 9B: The DNA sequence (SEQ ID NO: 9) of a portion of the human Notch insert is shown, starting near the 3' end, and proceeding in the 3' to 5' direction. The sequences shown are tentative, subject to confirmation by determination of overlapping sequences. Figure 10. DNA (SEQ ID NO: 10) and Amino Acid (SEQ ID NO: 11) Sequences of Human Notch Contained in Plasmid cDNA Clone hN3k. Figure 11. DNA (SEQ ID NO: 12) and Amino Acid (SEQ ID NO: 13) Sequences of Human Notch Contained in Plasmid cDNA Clone hN5k. Figure 12. Comparison of hN5k With Other Notch Homologs. Figure 12A. Schematic representation oi Drosophila Notch. Indicated are the signal sequence (signal), the 36 EGF-like repeats, the three Notch/liη-12 repeats, the transmembrane domain (TM), the six CDC 10 repeats, the OPA repeat, and the PEST (proline, glutamic acid, serine, threonine)-rich region. Figure 12B. Alignment of the deduced amino acid sequence of hN5k with sequences of other Notch homologs. Amino acids are numbered on the left side. The cdclO and PEST-rich regions are both boxed, and individual cdclO repeats are marked. Amino acids which are identical in three or more sequences are highlighted. The primers used to clone hN5k are indicated below the sequences from which they were designed. The nuclear localization sequence (NLS), casein kinase II (CKII), and cdc2 kinase (cdc2) sites of the putative CcN motif of the vertebrate Notch homologs are boxed. The possible bipartite nuclear targeting sequence (BNTS) and proximal phosphorylation sites oi Drosophila Notch are also boxed. Figure 13. Aligned amino acid sequences of Notch proteins of various species. humN: the human Notch protein encoded by the hN homolog (contained in part in plasmid hN5k) (SEQ ID NO: 19). TAN-1: the human Notch protein encoded by the TAN-1 homolog (SEQ ID NO:20) (the sequence shown is derived partly from our own work and partly from the TAN-1 sequence as published by Ellisen et al., 1991, Cell 66:649-661); Xen N: Xenopus Notch protein (Coffman et al., 1990, Science 249:1438-1441). Dros N: Drosophila Notch protein (Wharton et al., 1985, Cell 43:567-581). Structural domains are indicated. Figure 14. Immunocytochemical staining of breast cancer tissue from a human patient. Malignant breast tissue in a sample obtained from a human patient was embedded in a paraffin section, and subjected to immunocytochemical staining with anti-human Notch monoclonal antibody P4, directed against the TAN-1 protein. Non-malignant breast tissue exhibited much less staining (not shown). Figure 15. Immunocytochemical staining of colon tissue from a human patient with colon cancer. A colon tissue sample obtained from a patient with colon cancer was embedded in a paraffin section, and subjected to immunocytochemical staining with anti-human Notch monoclonal antibody PI , directed against the hN-encoded protein. Areas of increased staining are those areas in which malignant cells are present, as determined by cell morphology. Figure 16. Immunocytochemical staining of cervical tissue. Human tissue samples were obtained, containing cancer of the cervix (Fig. 16A) or normal cervical epithelium (Fig. 16B) from the same patient, embedded in a paraffin section, and subjected to immunocytochemical staining with anti-human Notch monoclonal antibody directed against the TAN-1 protein. Areas containing malignant cells (as determined by morphology) exhibited increasing staining relative to non-malignant cells. Among non-malignant cells, connective tissue and the basal layer of the epithelium (containing stem cells) stained with the anti- Notch antibody. Figure 17. DNA (SEQ ID NO:21) and encoded amino acid sequence (contained in SEQ ID NO: 19) of human Notch homolog hN. The entire DNA coding sequence is presented (as well as noncoding sequence), with the exclusion of that encoding the initiator Met. The last 8 nucleotides shown (numbers 9716-9723) are vector, and not hN, sequences. 5. DETAILED DESCRIPTION OF THE INVENTION The present invention relates to therapeutic and diagnostic methods and compositions based on Notch proteins and nucleic acids. The invention provides for treatment of disorders of cell fate or differentiation by administration of a therapeutic compound of the invention. Such therapeutic compounds (termed herein "Therapeutics") include: Notch proteins and analogs and derivatives (including fragments) thereof; antibodies thereto; nucleic acids encoding the Notch proteins, analogs, or derivatives; Notch antisense nucleic acids; as well as toporythmic proteins and derivatives and analogs thereof which bind to or otherwise interact with Notch proteins, and their encoding nucleic acids and antibodies. Also included are proteins and derivatives and analogs thereof which are capable of inhibiting the interactions of a Notch protein with another toporythmic protein (e.g. Delta, Serrate). In a preferred embodiment, a Therapeutic of the invention is administered to treat a cancerous condition, or to prevent progression from a pre-neoplastic or non-malignant state (e.g. , metaplastic condition) into a neoplastic or a malignant state. In another specific embodiment, a Therapeutic of the invention is administered to treat a nervous system disorder, such as nerve injury or a degenerative disease. In yet another specific embodiment, a Therapeutic of the invention is administered to promote tissue regeneration and repair for treatment of various conditions. In one embodiment, Therapeutics which antagonize, or inhibit, Notch function (hereinafter "Antagonist Therapeutics") are administered for therapeutic effect; disorders which can be thus treated can be identified by in vitro assays such as described in Section 5.1, infra. Such Antagonist Therapeutics include but are not limited to Notch antisense nucleic acids, anti-Notch neutralizing antibodies, competitive inhibitors of Notch protein-protein interactions (e.g. , a protein comprising Notch ELR-11 and ELR-12), and molecules which interfere with notch intracellular function such as that mediated by the cdclO repeats, as detailed infra. In another embodiment, Therapeutics which promote Notch function (hereinafter "Agonist Therapeutics") are administered for therapeutic effect; disorders which can thus be treated can be identified by in vitro assays such as described in Section 5.1, infra. Such Agonist Therapeutics include but are not limited to Notch proteins and derivatives thereof comprising the intracellular domain, Notch nucleic acids encoding the foregoing, and proteins comprising toporythmic protein domains that interact with Notch (e.g. , a protein comprising an extracellular domain of a Delta protein or a Delta sequence homologous to Drosophila Delta amino acids 1-230 (see Figure 1 and SEQ ID NO:2), or comprising a Serrate sequence homologous to Drosophila Serrate amino acids 79-282 (see Figure 5 and SEQ ID NO:4)). Disorders of cell fate, in particular precancerous conditions such as metaplasia and dysplasia, and hyperproliferative (e.g. , cancer) or hypoproliferative disorders, involving aberrant or undesirable levels of expression or activity of Notch protein can be diagnosed by detecting such levels, as described more fully infra. In a preferred aspect, a Therapeutic of the invention is a protein consisting of at least a fragment (termed herein "adhesive fragment") of the proteins encoded by toporythmic genes which mediates binding to Notch proteins or adhesive fragments thereof. Toporythmic genes, as used herein, shall mean the genes Notch. Delta, and Serrate, as well as other members of the Delta/Serrate family which may be identified by virtue of sequence homology or genetic interaction, and, more generally, members of the "Notch cascade" or the "Notch group" of genes, which are identified by molecular interactions (e.g. , binding in vitro) or genetic interactions (as detected phenotypically, e.g. , in Drosophila). The invention further provides a human Notch protein encoded by the hN homolog, and proteins comprising the extracellular domain of the Notch protein and subsequences thereof. Nucleic acids encoding the foregoing, and recombinant cells are also provided. For clarity of disclosure, and not by way of limitation, the detailed description of the invention is divided into the following subsections: (i) Therapeutic Uses; (ii) Prophylactic Uses; (iϋ) Demonstration of Therapeutic or Prophylactic Utility; (iv) Therapeutic/Prophylactic Administration and Compositions; (v) Antisense Regulation of Notch Expression; (vi) Diagnostic Utility; (vii) Notch Nucleic Acids; (viii) Recombinant Production of Protein Therapeutics; (ix) Derivatives and Analogs of Notch and Other Toporythmic Proteins; (x) Assays of Notch Proteins. Derivatives and Analogs; and (xi) Antibodies to Notch Proteins, Derivatives and Analogs. 5.1. THERAPEUTIC USES As stated supra, the Antagonist Therapeutics of the invention are those Therapeutics which antagonize, or inhibit, a Notch function. Such Antagonist Therapeutics are most preferably identified by use of known convenient in vitro assays, e.g. , based on their ability to inhibit binding of Notch to other proteins (see Sections 6-8 herein), or inhibit any known Notch function as assayed in vitro, although genetic assays (e.g. , in Drosophila) may also be employed. In a preferred embodiment, the Antagonist Therapeutic is a protein or derivative thereof comprising a functionally active fragment such as an adhesive fragment of Notch. In specific embodiments, such an Antagonist Therapeutic may be those adhesive proteins encoded by the appropriate constructs described in Sections 6 and 7 infra, or proteins comprising the Notch extracellular region, in particular ELR-11 and ELR-12, or an antibody thereto, or an analog/competitive inhibitor of a Notch intracellular signal-transducing region, a nucleic acid capable of expressing a Notch adhesive fragment, or a Notch antisense nucleic acid (see Section 5.5 herein). It should be noted that in certain instances, a Notch adhesive fragment (or possibly other presumed Antagonist Therapeutics) may alternatively act as an Agonist Therapeutic, depending on the developmental history of the tissue being exposed to the Therapeutic; preferably, suitable in vitro or in vivo assays, as described infra, should be utilized to determine the effect of a specific Therapeutic and whether its administration is indicated for treatment of the affected tissue. In another embodiment of the invention, a nucleic acid containing a portion of a Notch gene is used, as an Antagonist Therapeutic, to promote Notch inactivation by homologous recombination (Koller and Smithies, 1989, Proc. Natl. Acad. Sci. USA 86:8932-8935; Zijlstra et al., 1989, Nature 342:435- 438). The Agonist Therapeutics of the invention, as described supra, promote Notch function. Such Agonist Therapeutics include but are not limited to proteins and derivatives comprising the portions of toporythmic proteins such as Delta or Serrate that mediate binding to Notch, and nucleic acids encoding the foregoing (which can be administered to express their encoded products in vivo). In a specific embodiment, such a portion of Delta is D. melanogaster Delta amino acids 1-230 (SEQ ID NO: l) or a portion of a human Delta most homologous thereto. In another specific embodiment, such a portion of Serrate is D. melanogaster Serrate amino acids 79-282 (SEQ ID NO:5), or a portion of a human Serrate most homologous thereto. In other specific embodiments, such a portion of Delta or Serrate is the extracellular portion of such protein. Further descriptions and sources of Therapeutics of the inventions are found in Sections 5.4 through 5.8 herein. The Agonist and Antagonist Therapeutics of the invention have therapeutic utility for disorders of cell fate. The Agonist Therapeutics are administered therapeutically (including prophylactically): (1) in diseases or disorders involving an absence or decreased (relative to normal, or desired) levels of Notch function, for example, in patients where Notch protein is lacking, genetically defective, biologically inactive or underactive, or underexpressed; and (2) in diseases or disorders wherein in vitro (or in vivo) assays (see infra) indicate the utility of Notch agonist administration. The absence or decreased levels in Notch function can be readily detected, e.g. , by obtaining a patient tissue sample (e.g. , from biopsy tissue) and assaying it in vitro for protein levels, structure and/or activity of the expressed Notch protein. Many methods standard in the art can be thus employed, including but not limited to immunoassays to detect and/or visualize Notch protein (e.g. , Western blot, immunoprecipitation followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, immunocytochemistry, etc.; see also those assays listed in Section 5.6, infra), and/or hybridization assays to detect Notch expression by detecting and/or visualizing Notch mRNA (e.g. , Northern assays, dot blots, in situ hybridization, etc.) In vitro assays which can be used to determine whether administration of a specific Agonist Therapeutic or Antagonist Therapeutic is indicated, include in vitro cell culture assays in which a patient tissue sample is grown in culture, and exposed to or otherwise administered a Therapeutic, and the effect of such Therapeutic upon the tissue sample is observed. In one embodiment, where the patient has a malignancy, a sample of cells from such malignancy is plated out or grown in culture, and the cells are then exposed to a Therapeutic. A Therapeutic which inhibits survival or growth of the malignant cells (e.g. , by promoting terminal differentiation) is selected for therapeutic use in vivo. Many assays standard in the art can be used to assess such survival and/or growth; for example, cell proliferation can be assayed by measuring 3 H-thymidine incorporation, by direct cell count, by detecting changes in transcriptional activity of known genes such as proto-oncogenes (e.g. ,fos, myc) or cell cycle markers; cell viability can be assessed by trypan blue staining, differentiation can be assessed visually based on changes in morphology, etc. In a specific aspect, the malignant cell cultures are separately exposed to (1) an Agonist Therapeutic, and (2) an Antagonist Therapeutic; the result of the assay can indicate which type of Therapeutic has therapeutic efficacy. In another embodiment, a Therapeutic is indicated for use which exhibits the desired effect, inhibition or promotion of cell growth, upon a patient cell sample from tissue having or suspected of having a hyper- or hypoproliferative disorder, respectively. Such hyper- or hypoproliferative disorders include but are not limited to those described in Sections 5.1.1 through 5.1.3 infra. In another specific embodiment, a Therapeutic is indicated for use in treating nerve injury or a nervous system degenerative disorder (see Section 5.1.2) which exhibits in vitro promotion of nerve regeneration/neurite extension from nerve cells of the affected patient type. In addition, administration of an Antagonist Therapeutic of the invention is also indicated in diseases or disorders determined or known to involve a Notch dominant activated phenotype ("gain of function" mutations.) Administration of an Agonist Therapeutic is indicated in diseases or disorders determined or known to involve a Notch dominant negative phenotype ("loss of function" mutations). We have investigated the functions of various structural domains of the Notch protein in vivo, by ectopically expressing a series of Drosophila Notch deletion mutants under the hsp70 heat-shock promoter, as well as eye-specific promoters. Two classes of dominant phenotypes were observed, one suggestive of Notch loss-of function mutations and the other of Notch gain-of- function mutations. Dominant "activated" phenotypes resulted from overexpression of a protein lacking most extracellular sequences, while dominant "negative" phenotypes resulted from overexpression of a protein lacking most intracellular sequences. Our results indicate that Notch functions as a receptor whose extracellular domain mediates ligand-binding, resulting in the transmission of developmental signals by the cytoplasmic domain. The phenotypes observed also suggested that the cdclO/ankyrin repeat region within the intracellular domain plays an essential role in Notch mediated signal transduction events (intracellular function). In various specific embodiments, in vitro assays can be carried out with representative cells of cell types involved in a patient's disorder, to determine if a Therapeutic has a desired effect upon such cell types. In another embodiment, cells of a patient tissue sample suspected of being pre-neoplastic are similarly plated out or grown in vitro, and exposed to a Therapeutic. The Therapeutic which results in a cell phenotype that is more normal (i.e., less representative of a pre-neoplastic state, neoplastic state, malignant state, or transformed phenotype) is selected for therapeutic use. Many assays standard in the art can be used to assess whether a pre-neoplastic state, neoplastic state, or a transformed or malignant phenotype, is present (see Section 5.2.1). For example, characteristics associated with a transformed phenotype (a set of in vitro characteristics associated with a tumorigenic ability in vivo) include a more rounded cell morphology, looser substratum attachment, loss of contact inhibition, loss of anchorage dependence, release of proteases such as plasminogen activator, increased sugar transport, decreased serum requirement, expression of fetal antigens, disappearance of the 250.000 dalton surface protein, etc. (see Luria et al., 1978, General Virology, 3d Ed., John Wiley & Sons, New York pp. 436-446). in other specific embodiments, the in vitro assays described supra can be carried out using a cell line, rather than a cell sample derived from the specific patient to be treated, in which the cell line is derived from or displays characteristic(s) associated with the malignant, neoplastic or pre-neoplastic disorder desired to be treated or prevented, or is derived from the neural or other cell type upon which an effect is desired, according to the present invention. The Antagonist Therapeutics are administered therapeutical ly (including prophylactically): (1) in diseases or disorders involving increased (relative to normal, or desired) levels of Notch function, for example, where the Notch protein is overexpressed or overactive; and (2) in diseases or disorders wherein in vitro (or in vivo) assays indicate the utility of Notch antagonist administration. The increased levels of Notch function can be readily detected by methods such as those described above, by quantifying protein and/or RNA. In vitro assays with cells of patient tissue sample or the appropriate cell line or cell type, to determine therapeutic utility, can be carried out as described above. 5.1.1. MALIGNANCIES Malignant and pre-neoplastic conditions which can be tested as described supra for efficacy of intervention with Antagonist or Agonist Therapeutics, and which can be treated upon thus observing an indication of therapeutic utility, include but are not limited to those described below in Sections 5.1.1 and 5.2.1. Malignancies and related disorders, cells of which type can be tested in vitro (and/or in vivo), and upon observing the appropriate assay result, treated according to the present invention, include but are not limited to those listed in Table 1 (for a review of such disorders, see Fishman et al.. 1985, Medicine, 2d Ed., J.B. Lippincott Co., Philadelphia): TABLE 1 MALIGNANCIES AND RELATED DISORDERS Leukemia acute leukemia acute lymphocytic leukemia acute myelocytic leukemia myeloblastic promyelocytic myelomonocytic monocytic erythroleukemia chronic leukemia chronic myelocytic (granulocytic) leukemia chronic lymphocytic leukemia Polycythemia vera Lymphoma Hodgkin's disease non-Hodgkin's disease Multiple myeloma Waldenstrόm's macroglobulinemia Heavy chain disease Solid tumors sarcomas and carcinomas fibrosarcoma myxosarcoma liposarcoma chondrosarcoma osteogenic sarcoma chordoma angiosarcoma endotheliosarcoma lymphangiosarcoma lymphangioendotheliosarcoma synovioma mesothelioma Ewing's tumor leiomyosarcoma rhabdomyosarcoma colon carcinoma pancreatic cancer breast cancer ovarian cancer prostate cancer squamous cell carcinoma basal cell carcinoma adenocarcinoma sweat gland carcinoma sebaceous gland carcinoma papillary carcinoma papillary adenocarcinomas cystadenocarcinoma medullary carcinoma bronchogenic carcinoma renal cell carcinoma hepatoma bile duct carcinoma choriocarcinoma seminoma embryonal carcinoma Wilms' tumor cervical cancer testicular tumor lung carcinoma small cell lung carcinoma bladder carcinoma epithelial carcinoma glioma astrocytoma medulloblastoma craniopharyngioma ependymoma pinealoma hemangioblastoma acoustic neuroma oligodendroglioma menangioma melanoma neuroblastoma retinoblastoma In specific embodiments, malignancy or dysproliferative changes (such as metaplasias and dysplasias) are treated or prevented in epithelial tissues such as those in the cervix, esophagus, and lung. As detailed in the examples section 10.1 infra, malignancies of the breast, colon, and cervix exhibit increased expression of human Notch relative to such non-malignant tissue. Thus, in specific embodiments, malignancies of the breast, colon, or cervix are treated or prevented by administering an effective amount of an Antagonist Therapeutic of the invention. The presence of increased Notch expression in breast, colon, and cervical cancer suggests that many more cancerous conditions exhibit upregulated Notch. Thus, we envision that many more cancers, e.g. , seminoma, melanoma, and lung cancer, can be treated or prevented by administration of an Antagonist Therapeutic. 5.1.2. NERVOUS SYSTEM DISORDERS Nervous system disorders, involving cell types which can be tested as described supra for efficacy of intervention with Antagonist or Agonist Therapeutics, and which can be treated upon thus observing an indication of therapeutic utility, include but are not limited to nervous system injuries, and diseases or disorders which result in either a disconnection of axons, a diminution or degeneration of neurons, or demyelination. Nervous system lesions which may be treated in a patient (including human and non-human mammalian patients) according to the invention include but are not limited to the following lesions of either the central (including spinal cord, brain) or peripheral nervous systems: (i) traumatic lesions, including lesions caused by physical injury or associated with surgery, for example, lesions which sever a portion of the nervous system, or compression injuries; (ii) ischemic lesions, in which a lack of oxygen in a portion of the nervous system results in neuronal injury or death, including cerebral infarction or ischemia, or spinal cord infarction or ischemia; (iii) malignant lesions, in which a portion of the nervous system is destroyed or injured by malignant tissue which is either a nervous system associated malignancy or a malignancy derived from non-nervous system tissue; (iv) infectious lesions, in which a portion of the nervous system is destroyed or injured as a result of infection, for example, by an abscess or associated with infection by human immunodeficiency virus, herpes zoster, or herpes simplex virus or with Lyme disease, tuberculosis, syphilis; (v) degenerative lesions, in which a portion of the nervous system is destroyed or injured as a result of a degenerative process including but not limited to degeneration associated with Parkinson's disease, Alzheimer's disease, Huntington's chorea, or amyotrophic lateral sclerosis; (vi) lesions associated with nutritional diseases or disorders, in which a portion of the nervous system is destroyed or injured by a nutritional disorder or disorder of metabolism including but not limited to, vitamin B12 deficiency, folic acid deficiency, Wernicke disease, tobacco-alcohol amblyopia, Marchiafava-Bignami disease (primary degeneration of the corpus callosum), and alcoholic cerebellar degeneration; (vii) neurological lesions associated with systemic diseases including but not limited to diabetes (diabetic neuropathy, Bell's palsy), systemic lupus erythematosus, carcinoma, or sarcoidosis; (viii) lesions caused by toxic substances including alcohol, lead, or particular neurotoxins; and (ix) demyelinated lesions in which a portion of the nervous system is destroyed or injured by a demyelinating disease including but not limited to multiple sclerosis, human immunodeficiency virus-associated myelopathy, transverse myelopathy or various etiologies, progressive multifocal leukoencephalopathy, and central pontine myelinolysis. Therapeutics which are useful according to the invention for treatment of a nervous system disorder may be selected by testing for biological activity in promoting the survival or differentiation of neurons (see also Section 5.1). For example, and not by way of limitation, Therapeutics which elicit any of the following effects may be useful according to the invention: (i) increased survival time of neurons in culture; (ii) increased sprouting of neurons in culture or in vivo; (iii) increased production of a neuron-associated molecule in culture or in vivo, e.g. , choline acetyltransferase or acetylcholinesterase with respect to motor neurons; or (iv) decreased symptoms of neuron dysfunction in vivo. Such effects may be measured by any method known in the art. In preferred, non-limiting embodiments, increased survival of neurons may be measured by the method set forth in Arakawa et al. (1990, J. Neurosci. 10:3507-3515); increased sprouting of neurons may be detected by methods set forth in Pestronk et al. (1980, Exp. Neurol. 70:65-82) or Brown et al. (1981 , Ann. Rev. Neurosci. 4:17-42); increased production of neuron-associated molecules may be measured by bioassay, enzymatic assay, antibody binding, Northern blot assay, etc., depending on the molecule to be measured; and motor neuron dysfunction may be measured by assessing the physical manifestation of motor neuron disorder, e.g. , weakness, motor neuron conduction velocity, or functional disability. In a specific embodiments, motor neuron disorders that may be treated according to the invention include but are not limited to disorders such as infarction, infection, exposure to toxin, trauma, surgical damage, degenerative disease or malignancy that may affect motor neurons as well as other components of the nervous system, as well as disorders that selectively affect neurons such as amyotrophic lateral sclerosis, and including but not limited to progressive spinal muscular atrophy, progressive bulbar palsy, primary lateral sclerosis, infantile and juvenile muscular atrophy, progressive bulbar paralysis of childhood (Fazio- Londe syndrome), poliomyelitis and the post polio syndrome, and Hereditary Motorsensory Neuropathy (Charcot-Marie-Tooth Disease). 5.1.3. TISSUE REPAIR AND REGENERATION In another embodiment of the invention, a Therapeutic of the invention is used for promotion of tissue regeneration and repair, including but not limited to treatment of benign dysproliferative disorders. Specific embodiments are directed to treatment of cirrhosis of the liver (a condition in which scarring has overtaken normal liver regeneration processes), treatment of keloid (hypertrophic scar) formation (disfiguring of the skin in which the scarring process interferes with normal renewal), psoriasis (a common skin condition characterized by excessive proliferation of the skin and delay in proper cell fate determination), and baldness (a condition in which terminally differentiated hair follicles (a tissue rich in Notch) fail to function properly). 5.2. PROPHYLACTIC USES 5.2.1. MALIGNANCIES The Therapeutics of the invention can be administered to prevent progression to a neoplastic or malignant state, including but not limited to those disorders listed in Table 1. Such administration is indicated where the Therapeutic is shown in assays, as described supra, to have utility for treatment or prevention of such disorder. Such prophylactic use is indicated in conditions known or suspected of preceding progression to neoplasia or cancer, in particular, where non-neoplastic cell growth consisting of hypeφlasia, metaplasia, or most particularly, dysplasia has occurred (for review of such abnormal growth conditions, see Robbins and Angell, 1976, Basic Pathology, 2d Ed., W.B. Saunders Co., Philadelphia, pp. 68-79.) Hypeφlasia is a form of controlled cell proliferation involving an increase in cell number in a tissue or organ, without significant alteration in structure or function. As but one example, endometrial hypeφlasia often precedes endometrial cancer. Metaplasia is a form of controlled cell growth in which one type of adult or fully differentiated cell substitutes for another type of adult cell. Metaplasia can occur in epithelial or connective tissue cells. Atypical metaplasia involves a somewhat disorderly metaplastic epithelium. Dysplasia is frequently a forerunner of cancer, and is found mainly in the epithelia; it is the most disorderly form of non-neoplastic cell growth, involving a loss in individual cell uniformity and in the architectural orientation of cells. Dysplastic cells often have abnormally large, deeply stained nuclei, and exhibit pleomoφhism. Dysplasia characteristically occurs where there exists chronic irritation or inflammation, and is often found in the cervix, respiratory passages, oral cavity, and gall bladder. Alternatively or in addition to the presence of abnormal cell growth characterized as hypeφlasia, metaplasia, or dysplasia, the presence of one or more characteristics of a transformed phenotype, or of a malignant phenotype, displayed in vivo or displayed in vitro by a cell sample from a patient, can indicate the desirability of prophylactic/therapeutic administration of a Therapeutic of the invention. As mentioned supra, such characteristics of a transformed phenotype include moφhology changes, looser substratum attachment, loss of contact inhibition, loss of anchorage dependence, protease release, increased sugar transport, decreased serum requirement, expression of fetal antigens, disappearance of the 250,000 dalton cell surface protein, etc. (see also id., at pp. 84-90 for characteristics associated with a transformed or malignant phenotype). In a specific embodiment, leukoplakia, a benign-appearing hypeφlastic or dysplastic lesion of the epithelium, or Bowen's disease, a carcinoma in situ, are pre-neoplastic lesions indicative of the desirability of prophylactic intervention. In another embodiment, fibrocystic disease (cystic hypeφlasia, mammary dysplasia, particularly adenosis (benign epithelial hypeφlasia)) is indicative of the desirability of prophylactic intervention. In other embodiments, a patient which exhibits one or more of the following predisposing factors for malignancy is treated by administration of an effective amount of a Therapeutic: a chromosomal translocation associated with a malignancy (e.g. , the Philadelphia chromosome for chronic myelogenous leukemia, t(14;18) for follicular lymphoma, etc.), familial polyposis or Gardner's syndrome (possible forerunners of colon cancer), benign monoclonal gammopathy (a possible forerunner of multiple myeloma), and a first degree kinship with persons having a cancer or precancerous disease showing a Mendelian (genetic) inheritance pattern (e.g. , familial polyposis of the colon, Gardner's syndrome, hereditary exostosis, polyendocrine adenomatosis, medullary thyroid carcinoma with amyloid production and pheochromocytoma, Peutz-Jeghers syndrome, neurofibromatosis of Von Recklinghausen, retinoblastoma, carotid body tumor, cutaneous melanocarcinoma, intraocular melanocarcinoma, xeroderma pigmentosum, ataxia telangiectasia, Chediak-Higashi syndrome, albinism, Fanconi's aplastic anemia, and Bloom's syndrome; see Robbins and Angell, 1976, Basic Pathology, 2d Ed., W.B. Saunders Co., Philadelphia, pp. 112-113) etc.) In another specific embodiment, an Antagonist Therapeutic of the invention is administered to a human patient to prevent progression to breast, colon, or cervical cancer. 5.2.2. OTHER DISORDERS In other embodiments, a Therapeutic of the invention can be 0 administered to prevent a nervous system disorder described in Section 5.1.2, or other disorder (e.g. , liver cirrhosis, psoriasis, keloids, baldness) described in Section 5.1.3. 5 5.3. DEMONSTRATION OF THERAPEUTIC OR PROPHYLACTIC UTILITY The Therapeutics of the invention can be tested in vivo for the desired therapeutic or prophylactic activity. For example, such compounds can be tested in suitable animal model systems prior to testing in humans, including o but not limited to rats, mice, chicken, cows, monkeys, rabbits, etc. For in vivo testing, prior to administration to humans, any animal model system known in the art may be used. 5.4. THERAPEUTIC/PROPHYLACTIC ADMINISTRATION AND COMPOSITIONS 5 The invention provides methods of treatment (and prophylaxis) by administration to a subject of an effective amount of a Therapeutic of the invention. In a preferred aspect, the Therapeutic is substantially purified. The subject is preferably an animal, including but not limited to animals such as cows, pigs, chickens, etc., and is preferably a mammal, and most preferably human. 0 Various delivery systems are known and can be used to administer a Therapeutic of the invention, e.g. , encapsulation in liposomes, microparticles, 5 microcapsules, expression by recombinant cells, receptor-mediated endocytosis (see, e.g. , Wu and Wu, 1987, J. Biol. Chem. 262:4429-4432), construction of a Therapeutic nucleic acid as part of a retroviral or other vector, etc. Methods of introduction include but are not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, and oral routes. The compounds may be administered by any convenient route, for example by infusion or bolus injection, by absoφtion through epithelial or mucocutaneous linings (e.g. , oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other biologically active agents. Administration can be systemic or local. In addition, it may be desirable to introduce the pharmaceutical compositions of the invention into the central nervous system by any suitable route, including intraventricular and intrathecal injection; intraventricular injection may be facilitated by an intraventricular catheter, for example, attached to a reservoir, such as an Om aya reservoir. In a specific embodiment, it may be desirable to administer the pharmaceutical compositions of the invention locally to the area in need of treatment; this may be achieved by, for example, and not by way of limitation, local infusion during surgery, topical application, e.g. , in conjunction with a wound dressing after surgery, by injection, by means of a catheter, by means of a suppository, or by means of an implant, said implant being of a porous, non- porous, or gelatinous material, including membranes, such as sialastic membranes, or fibers. In one embodiment, administration can be by direct injection at the site (or former site) of a malignant tumor or neoplastic or pre- neoplastic tissue. in a specific embodiment, administration of a Therapeutic into a Notch-expressing cell is accomplished by linkage of the Therapeutic to a Delta (or other toporythmic) protein or portion thereof capable of mediating binding to Notch. Contact of a Notch-expressing cell with the linked Therapeutic results in binding of the linked Therapeutic via its Delta portion to Notch on the surface of the cell, followed by uptake of the linked Therapeutic into the Notch-expressing cell. In a specific embodiment wherein an analog of a Notch intracellular signal-transducing domain is employed as a Therapeutic, such that it can inhibit Notch signal transduction, the analog is preferably delivered intracellularly (e.g. , by expression from a nucleic acid vector, or by linkage to a Delta protein capable of binding to Notch followed by binding and internalization, or by receptor-mediated mechanisms). In a specific embodiment where the Therapeutic is a nucleic acid encoding a protein Therapeutic, the nucleic acid can be administered in vivo to promote expression of its encoded protein, by constructing it as part of an appropriate nucleic acid expression vector and administering it so that it becomes intracellular, e.g. , by use of a retroviral vector (see U.S. Patent No. 4,980,286), or by direct injection, or by use of microparticle bombardment (e.g. , a gene gun; Biolistic, Dupont), or coating with lipids or cell-surface receptors or transfecting agents, or by administering it in linkage to a homeobox-like peptide which is known to enter the nucleus (see e.g. , Joliot et al., 1991 , Proc. Natl. Acad. Sci. USA 88: 1864-1868), etc. Alternatively, a nucleic acid Therapeutic can be introduced intracellularly and incoφorated within host cell DNA for expression, by homologous recombination. In specific embodiments directed to treatment or prevention of particular disorders, preferably the following forms of administration are used: Disorder Preferred Forms of Administration Cervical cancer Topical Gastrointestinal cancer Oral; intravenous Lung cancer Inhaled; intravenous Leukemia Intravenous; extracoφoreal Metastatic carcinomas Intravenous; oral Brain cancer Targeted; intravenous; intrathecal Liver cirrhosis Oral; intravenous Psoriasis Topical Keloids Topical Baldness Topical Spinal cord injury Targeted; intravenous; intrathecal Parkinson's disease Targeted; intravenous; intrathecal Motor neuron disease Targeted; intravenous; intrathecal Alzheimer's disease Targeted; intravenous; intrathecal The present invention also provides pharmaceutical compositions. Such compositions comprise a therapeutical ly effective amount of a Therapeutic, and a pharmaceutically acceptable carrier or excipient. Such a carrier includes but is not limited to saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof. The carrier and composition can be sterile. The formulation should suit the mode of administration. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. The composition can be a liquid solution, suspension, emulsion, tablet, pill, capsule, sustained release formulation, or powder. The composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides. Oral formulation can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. In a preferred embodiment, the composition is formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous administration to human beings. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. Where necessary, the composition may also include a solubilizing agent and a local anesthetic such as lignocaine to ease pain at the site of the injection. Generally, the ingredients are supplied either separately or mixed together in unit dosage form, for example, as a dry lyophilized powder or water free concentrate in a hermetically sealed container such as an ampoule or sachette indicating the quantity of active agent. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients may be mixed prior to administration. The Therapeutics of the invention can be formulated as neutral or salt forms. Pharmaceutically acceptable salts include those formed with free amino groups such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with free carboxyl groups such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc. The amount of the Therapeutic of the invention which will be effective in the treatment of a particular disorder or condition will depend on the nature of the disorder or condition, and can be determined by standard clinical techniques. In addition, in vitro assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the seriousness of the disease or disorder, and should be decided according to the judgment of the practitioner and each patient's circumstances. However, suitable dosage ranges for intravenous administration are generally about 20-500 micrograms of active compound per kilogram body weight. Suitable dosage ranges for intranasal administration are generally about 0.01 pg/kg body weight to 1 mg/kg body weight. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems. Suppositories generally contain active ingredient in the range of 0.5% to 10% by weight; oral formulations preferably contain 10% to 95% active ingredient. The invention also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the invention. Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration. 5.5. ANTISENSE REGULATION OF NOTCH EXPRESSION The present invention provides the therapeutic or prophylactic use of nucleic acids of at least six nucleotides that are antisense to a gene or cDNA encoding Notch or a portion thereof. "Antisense" as used herein refers to a nucleic acid capable of hybridizing to a portion of a Notch RNA (preferably mRNA) by virtue of some sequence complementarity. Such antisense nucleic acids have utility as Antagonist Therapeutics of the invention, and can be used in the treatment or prevention of disorders as described supra in Section 5.1 and its subsections. The antisense nucleic acids of the invention can be oligonucleotides that are double-stranded or single-stranded, RNA or DNA or a modification or derivative thereof, which can be directly administered to a cell, or which can be produced intracellularly by transcription of exogenous, introduced sequences. In a specific embodiment, the Notch antisense nucleic acids provided by the instant invention can be used for the treatment of tumors or other disorders, the cells of which tumor type or disorder can be demonstrated (in vitro or in vivo) to express the Notch gene. Such demonstration can be by detection of Notch RNA or of Notch protein. The invention further provides pharmaceutical compositions comprising an effective amount of the Notch antisense nucleic acids of the invention in a pharmaceutically acceptable carrier, as described supra in Section 5.4. Methods for treatment and prevention of disorders (such as those described in Sections 5.1 and 5.2) comprising administering the pharmaceutical compositions of the invention are also provided. In another embodiment, the invention is directed to methods for inhibiting the expression of a Notch nucleic acid sequence in a prokaryotic or eukaryotic cell comprising providing the cell with an effective amount of a composition comprising an antisense Notch nucleic acid of the invention. In another embodiment, the identification of cells expressing functional Notch receptors can be carried out by observing the ability of Notch to "rescue" such cells from the cytotoxic effects of a Notch antisense nucleic acid. In an alternative embodiment of the invention, nucleic acids antisense to a nucleic acid encoding a ("adhesive") toporythmic protein or fragment that binds to Notch, are envisioned as Therapeutics. Notch antisense nucleic acids and their uses are described in detail below. 5.5.1. NOTCH ANTISENSE NUCLEIC ACIDS The Notch antisense nucleic acids are of at least six nucleotides and are preferably oligonucleotides (ranging from 6 to about 50 oligonucleotides). In specific aspects, the oligonucleotide is at least 10 nucleotides, at least 15 nucleotides, at least 100 nucleotides, or at least 200 nucleotides. The oligonucleotides can be DNA or RNA or chimeric mixtures or derivatives or modified versions thereof, single-stranded or double-stranded. The oligonucleotide can be modified at the base moiety, sugar moiety, or phosphate backbone. The oligonucleotide may include other appending groups such as peptides, or agents facilitating transport across the cell membrane (see, e.g. , Letsinger et al., 1989, Proc. Natl. Acad. Sci. U.S.A. 86:6553-6556; Lemaitre et al., 1987, Proc. Natl. Acad. Sci. 84:648-652; PCT Publication No. WO 88/09810, published December 15, 1988) or blood-brain barrier (see, e.g. , PCT Publication No. WO 89/10134, published April 25. 1988), hybridization- triggered cleavage agents (see, e.g. , Krol et al., 1988, BioTechniques 6:958-976) or intercalating agents (see, e.g. , Zon, 1988, Pharm. Res. 5:539-549). In a preferred aspect of the invention, a Notch antisense oligonucleotide is provided, preferably of single-stranded DNA. In a most preferred aspect, such an oligonucleotide comprises a sequence antisense to the sequence encoding ELR 11 and ELR 12 of Notch, most preferably, of human Notch. The oligonucleotide may be modified at any position on its structure with substituents generally known in the art. The Notch antisense oligonucleotide may comprise at least one modified base moiety which is selected from the group including but not limited to 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xantine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl- 2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2- carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. In another embodiment, the oligonucleotide comprises at least one modified sugar moiety selected from the group including but not limited to arabinose, 2-fluoroarabinose, xylulose, and hexose. In yet another embodiment, the oligonucleotide comprises at least one modified phosphate backbone selected from the group consisting of a phosphorothioate, a phosphorodithioate, a phosphoramidothioate, a phosphoramidate, a phosphordiamidate, a methylphosphonate, an alkyl phosphotriester, and a formacetal or analog thereof. In yet another embodiment, the oligonucleotide is an α-anomeric oligonucleotide. An α-anomeric oligonucleotide forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual /3-units, the strands run parallel to each other (Gautier et al. , 1987, Nucl. Acids Res. 15:6625-6641). The oligonucleotide may be conjugated to another molecule, e.g. , a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent, etc. Oligonucleotides of the invention may be synthesized by standard methods known in the art, e.g. by use of an automated DNA synthesizer (such as are commercially available from Biosearch, Applied Biosystems, etc.). As examples, phosphorothioate oligos may be synthesized by the method of Stein et al. (1988, Nucl. Acids Res. 16:3209), methylphosphonate oligos can be prepared by use of controlled pore glass polymer supports (Sarin et al., 1988, Proc. Natl. Acad. Sci. U.S.A. 85:7448-7451), etc. In a specific embodiment, the Notch antisense oligonucleotide comprises catalytic RNA, or a ribozyme (see, e.g. , PCT International Publication WO 90/11364, published October 4, 1990; Sarver et al., 1990, Science 247: 1222- 1225). In another embodiment, the oligonucleotide is a 2'-0-methylribonucleotide (Inoue et al., 1987, Nucl. Acids Res. 15:6131-6148), or a chimeric RNA-DNA analogue (Inoue et al., 1987, FEBS Lett. 215:327-330). In an alternative embodiment, the Notch antisense nucleic acid of the invention is produced intracellularly by transcription from an exogenous sequence. For example, a vector can be introduced in vivo such that it is taken up by a cell, within which cell the vector or a portion thereof is transcribed, producing an antisense nucleic acid (RNA) of the invention. Such a vector would contain a sequence encoding the Notch antisense nucleic acid. Such a vector can remain episomal or become chromosomally integrated, as long as it can be transcribed to produce the desired antisense RNA. Such vectors can be constructed by recombinant DNA technology methods standard in the art. Vectors can be plasmid, viral, or others known in the art, used for replication and expression in mammalian cells. Expression of the sequence encoding the Notch antisense RNA can be by any promoter known in the art to act in mammalian, preferably human, cells. Such promoters can be inducible or constitutive. Such promoters include but are not limited to: the SV40 early promoter region (Bernoist and Chambon, 1981, Nature 290:304-310), the promoter contained in the 3' long terminal repeat of Rous sarcoma virus (Yamamoto et al. , 1980, Cell 22:787-797), the heφes thymidine kinase promoter (Wagner et al. , 1981 , Proc. Natl. Acad. Sci. U.S.A. 78: 1441-1445), the regulatory sequences of the metallothionein gene (Brinster et al. , 1982, Nature 296:39-42), etc. The antisense nucleic acids of the invention comprise a sequence complementary to at least a portion of an RNA transcript of a Notch gene, preferably a human Notch gene. However, absolute complementarity, although preferred, is not required. A sequence "complementary to at least a portion of an RNA," as referred to herein, means a sequence having sufficient complementarity to be able to hybridize with the RNA, forming a stable duplex; in the case of 5 double-stranded Notch antisense nucleic acids, a single strand of the duplex DNA may thus be tested, or triplex formation may be assayed. The ability to hybridize will depend on both the degree of complementarity and the length of the antisense nucleic acid. Generally, the longer the hybridizing nucleic acid, the more base mismatches with a Notch RNA it may contain and still form a stable duplex (or 10 triplex, as the case may be). One skilled in the art can ascertain a tolerable degree of mismatch by use of standard procedures to determine the melting point of the hybridized complex. 5.5.2. THERAPEUTIC UTILITY OF NOTCH j 5 ANTISENSE NUCLEIC ACIDS The Notch antisense nucleic acids can be used to treat (or prevent) malignancies, of a cell type which has been shown to express Notch RNA. Malignant, neoplastic, and pre-neoplastic cells which can be tested for such expression include but are not limited to those described supra in Sections 5.1.1 0 and 5.2.1. In a preferred embodiment, a single-stranded DNA antisense Notch oligonucleotide is used. Malignant (particularly, tumor) cell types which express Notch RNA can be identified by various methods known in the art. Such methods include but are not limited to hybridization with a Notch-specific nucleic acid 5 (e.g. by Northern hybridization, dot blot hybridization, in situ hybridization), observing the ability of RNA from the cell type to be translated in vitro into Notch, etc. In a preferred aspect, primary tumor tissue from a patient can be assayed for Notch expression prior to treatment. Pharmaceutical compositions of the invention (see Section 5.1.4), 30 comprising an effective amount of a Notch antisense nucleic acid in a pharmaceutically acceptable carrier, can be administered to a patient having a malignancy which is of a type that expresses Notch RNA. 35 The amount of Notch antisense nucleic acid which will be effective in the treatment of a particular disorder or condition will depend on the nature of the disorder or condition, and can be determined by standard clinical techniques. Where possible, it is desirable to determine the antisense cytotoxicity of the tumor type to be treated in vitro, and then in useful animal model systems prior to testing and use in humans. In a specific embodiment, pharmaceutical compositions comprising Notch antisense nucleic acids are administered via liposomes, microparticles, or microcapsules. In various embodiments of the invention, it may be useful to use such compositions to achieve sustained release of the Notch antisense nucleic acids. In a specific embodiment, it may be desirable to utilize liposomes targeted via antibodies to specific identifiable tumor antigens (Leonetti et al., 1990, Proc. Natl. Acad. Sci. U.S.A. 87:2448-2451; Renneisen et al.. 1990, J. Biol. Chem. 265: 16337-16342). 5.6. DIAGNOSTIC UTILITY Notch proteins, analogues, derivatives, and subsequences thereof, Notch nucleic acids (and sequences complementary thereto). anti-Notch antibodies, and other toporythmic proteins and derivatives and analogs thereof which interact with Notch proteins, and inhibitors of North-toporythmic protein interactions, have uses in diagnostics. Such molecules can be used in assays, such as immunoassays, to detect, prognose, diagnose, or monitor various conditions, diseases, and disorders affecting Notch expression, or monitor the treatment thereof. In particular, such an immunoassay is carried out by a method comprising contacting a sample derived from a patient with an anti-Notch antibody under conditions such that immunospecific binding can occur, and detecting or measuring the amount of any immunospecific binding by the antibody. In a specific embodiment, antibody to Notch can be used to assay in a patient tissue or serum sample for the presence of Notch where an aberrant level of Notch is an indication of a diseased condition. The immunoassays which can be used include but are not limited to competitive and non-competitive assay systems using techniques such as western blots, radioimmunoassays, ELISA (enzyme linked immunosorbent assay), "sandwich" immunoassays, immunoprecipitation assays, precipitin reactions, gel diffusion precipitin reactions, immunodiffusion assays, agglutination assays, complement-fixation assays, immunoradiometric assays, fluorescent immunoassays, protein A immunoassays, to name but a few. Notch genes and related nucleic acid sequences and subsequences, including complementary sequences, and other toporythmic gene sequences, can also be used in hybridization assays. Notch nucleic acid sequences, or subsequences thereof comprising about at least 8 nucleotides, can be used as hybridization probes. Hybridization assays can be used to detect, prognose, diagnose, or monitor conditions, disorders, or disease states associated with aberrant changes in Notch expression and/or activity as described supra. In particular, such a hybridization assay is carried out by a method comprising contacting a sample containing nucleic acid with a nucleic acid probe capable of hybridizing to Notch DNA or RNA, under conditions such that hybridization can occur, and detecting or measuring any resulting hybridization. As detailed in examples section 10.1 infra, increased Notch expression occurs in human breast, colon, and cervical cancer. Accordingly, in specific embodiments, human breast, colon, or cervical cancer or premalignant changes in such tissues is diagnosed by detecting increased Notch expression (or amount) in patient samples relative to the level of Notch expression (or amount) in an analogous non-malignant, or non-premalignant, as the case may be, sample (from the patient or another person, as determined experimentally or as is known as a standard level in such samples). In one embodiment, the Notch protein (or derivative having Notch antigenicity) that is detected or measured is on the cell surface. In another embodiment, the Notch protein (or derivative) is a cell free soluble molecule (e.g. , as measured in a blood or serum sample) or is intracellular. Without intending to be bound mechanistically, Applicants believe that cell free Notch may result from secretion or shedding from the cell surface. In yet another embodiment, soluble, cell-surface, and intracellular amounts of Notch protein or derivative are detected or measured. 5.7. NOTCH NUCLEIC ACIDS Therapeutics of the invention which are Notch nucleic acids or Notch antisense nucleic acids, as well as nucleic acids encoding protein Therapeutics, include those described below, which can be obtained by methods known in die art, and in particular, as described below. In particular aspects, the invention provides amino acid sequences of Notch, preferably human Notch, and fragments and derivatives thereof which comprise an antigenic determinant (i.e., can be recognized by an antibody) or which are functionally active, as well as nucleic acid sequences encoding die foregoing. "Functionally active" material as used herein refers to that material displaying one or more known functional activities associated with the full-length (wild-type) Notch protein product, e.g. , binding to Delta, binding to Serrate, binding to any other Notch ligand, antigenicity (binding to an anti-Notch antibody), etc. In specific embodiments, the invention provides fragments of a Notch protein consisting of at least 40 amino acids, or of at least 75 amino acids. In other embodiments, the proteins comprise or consist essentially of the intracellular domain, transmembrane region, extracellular domain, cdclO region, Notch/lin-12 repeats, or the EGF-homologous repeats, or any combination of the foregoing, of a Notch protein. Fragments, or proteins comprising fragments, lacking some or all of the EGF-homologous repeats of Notch are also provided. Nucleic acids encoding the foregoing are provided. In other specific embodiments, the invention provides nucleotide sequences and subsequences of Notch, preferably human Notch, consisting of at least 25 nucleotides, at least 50 nucleotides, or at least 150 nucleotides. Nucleic acids encoding the proteins and protein fragments described above are provided, as well as nucleic acids complementary to and capable of hybridizing to such nucleic acids. In one embodiment, such a complementary sequence may be complementary to a Notch cDNA sequence of at least 25 nucleotides, or of at least 100 nucleotides. In a preferred aspect, the invention utilizes cDNA sequences encoding human Notch or a portion thereof. In a specific embodiment, such sequences of the human Notch gene or cDNA are as contained in plasmids hN3k, hN4k, or hN5k (see Section 9, infra) or in the gene corresponding thereto; such a human Notch protein sequence can be as shown in Figures 10 (SEQ ID NO: 11) or 11 (SEQ ID NO: 13). In other embodiments, the Notch nucleic acid and/or its encoded protein has at least a portion of the sequence shown in one of the following publications: Wharton et al. , 1985, Cell 43:567-581 (Drosophila Notch); Kidd et al., 1986, Mol. Cell. Biol. 6:3094-3108 (Drosophila Notch); Coffman et al. , 1990, Science 249: 1438-1441 (Xenopus Notch); Ellisen et al., 1991 , Cell 66:649-661 (a human Notch). In another aspect, the sequences of human Notch are those encoding the human Notch amino acid sequences or a portion thereof as shown in Figure 13. In a particular aspect, the human Notch sequences are those of the hN homolog (represented in part by plasmid hN5k) or the TAN-1 homolog. In one embodiment of the invention, the invention is directed to the full-length human Notch protein encoded by the hN homolog as depicted in Figure 13, both containing the signal sequence (Le.. the precursor protein; amino acids 1-2169) and lacking the signal sequence (i.e. , the mature protein; amino acids ~26-2169), as well as portions of the foregoing (e.g. , the extracellular domain, EGF homologous repeat region, EGF-like repeats 11 and 12, cdc-10/ankyrin repeats, etc.) and proteins comprising the foregoing, as well as nucleic acids encoding the foregoing. As is readily apparent, as used herein, a "nucleic acid encoding a fragment or portion of a Notch protein" shall be construed as referring to a nucleic acid encoding only the recited fragment or portion of the Notch protein and not other portions of the Notch protein. In a preferred, but not limiting, aspect of the invention, a human Notch DNA sequence can be cloned and sequenced by the method described in Section 9, infra. In another preferred aspect, PCR is used to amplify the desired sequence in the library, prior to selection. For example, oligonucleotide primers representing part of the adhesive domains encoded by a homologue of the desired gene can be used as primers in PCR. The above-methods are not meant to limit the following general description of methods by which clones of Notch may be obtained. Any eukaryotic cell can potentially serve as the nucleic acid source for the molecular cloning of the Notch gene. The DNA may be obtained by standard procedures known in the art from cloned DNA (e.g.. a DNA "library"), by chemical synthesis, by cDNA cloning, or by the cloning of genomic DNA, or fragments thereof, purified from the desired human cell (see, for example Sambrook et al., 1989, Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory, 2d. Ed., Cold Spring Harbor, New York; Glover, D.M. (ed.), 1985, DNA Cloning: A Practical Approach, MRL Press, Ltd., Oxford, U.K. Vol. I, II.) Clones derived from genomic DNA may contain regulatory and intron DNA regions in addition to coding regions; clones derived from cDNA will contain only exon sequences. Whatever the source, the gene should be molecularly cloned into a suitable vector for propagation of the gene. In the molecular cloning of the gene from genomic DNA, DNA fragments are generated, some of which will encode the desired gene. The DNA may be cleaved at specific sites using various restriction enzymes. Alternatively, one may use DNAse in the presence of manganese to fragment the DNA, or the DNA can be physically sheared, as for example, by sonication. The linear DNA fragments can then be separated according to size by standard techniques, including but not limited to, agarose and polyacrylamide gel electrophoresis and column chromatography. Once the DNA fragments are generated, identification of the specific DNA fragment containing the desired gene may be accomplished in a number of ways. For example, if an amount of a portion of a Notch (of any species) gene or its specific RNA, or a fragment thereof e.g. , the adhesive domain, is available and can be purified and labeled, the generated DNA fragments may be screened by nucleic acid hybridization to the labeled probe (Benton, W. and Davis, R., 1977, Science 196, 180; Grunstein, M. And Hogness, D., 1975, Proc. Natl. Acad. Sci. U.S.A. 72, 3961). Those DNA fragments with substantial homology to the probe will hybridize. It is also possible to identify the appropriate fragment by restriction enzyme digestion(s) and comparison of fragment sizes with those expected according to a known restriction map if such is available. Further selection can be carried out on the basis of the properties of the gene. Alternatively, the presence of the gene may be detected by assays based on the physical, chemical, or immunological properties of its expressed product. For example, cDNA clones, or DNA clones which hybrid-select the proper mRNAs, can be selected which produce a protein that, e.g. , has similar or identical electrophoretic migration, isolectric focusing behavior, proteolytic digestion maps, in vitro aggregation activity ("adhesiveness") or antigenic properties as known for Notch. If an antibody to Notch is available, the Notch protein may be identified by binding of labeled antibody to the putatively Notch synthesizing clones, in an ELISA (enzyme-linked immunosorbent assay )-type procedure. The Notch gene can also be identified by mRNA selection by nucleic acid hybridization followed by in vitro translation. In this procedure, fragments are used to isolate complementary mRNAs by hybridization. Such DNA fragments may represent available, purified Notch DNA of another species (e.g. , Drosophila). Immunoprecipitation analysis or functional assays (e.g. , aggregation ability in vitro; see examples infra) of the in vitro translation products of the isolated products of the isolated mRNAs identifies the mRNA and, therefore, the complementary DNA fragments that contain the desired sequences. In addition, specific mRNAs may be selected by adsoφtion of polysomes isolated from cells to immobilized antibodies specifically directed against Notch or Delta protein. A radiolabelled Notch cDNA can be synthesized using the selected mRNA (from the adsorbed polysomes) as a template. The radiolabelled mRNA or cDNA may then be used as a probe to identify the Notch DNA fragments from among other genomic DNA fragments. Alternatives to isolating the Notch genomic DNA include, but are not limited to, chemically synthesizing the gene sequence itself from a known sequence or making cDNA to the mRNA which encodes the Notch gene. For example, RNA for cDNA cloning of the Notch gene can be isolated from cells which express Notch. Other methods are possible and within the scope of the invention. The identified and isolated gene can then be inserted into an appropriate cloning vector. A large number of vector-host systems known in the art may be used. Possible vectors include, but are not limited to, plasmids or modified viruses, but the vector system must be compatible with the host cell used. Such vectors include, but are not limited to, bacteriophages such as lambda derivatives, or plasmids such as PBR322 or pUC plasmid derivatives. The insertion into a cloning vector can, for example, be accomplished by ligating the DNA fragment into a cloning vector which has complementary cohesive termini. However, if the complementary restriction sites used to fragment the DNA are not present in the cloning vector, the ends of the DNA molecules may be enzymatically modified. Alternatively, any site desired may be produced by ligating nucleotide sequences (linkers) onto the DNA termini; these ligated linkers may comprise specific chemically synthesized oligonucleotides encoding restriction endonuclease recognition sequences. In an alternative method, the cleaved vector and Notch or Delta gene may be modified by homopolymeric tailing. Recombinant molecules can be introduced into host cells via transformation, transfection, infection, electroporation, etc. , so that many copies of the gene sequence are generated. In an alternative method, the desired gene may be identified and isolated after insertion into a suitable cloning vector in a "shot gun" approach. Enrichment for the desired gene, for example, by size fractionization, can be done before insertion into the cloning vector. In specific embodiments, transformation of host cells with recombinant DNA molecules that incoφorate the isolated Notch gene, cDNA, or synthesized DNA sequence enables generation of multiple copies of the gene. Thus, the gene may be obtained in large quantities by growing transformants, isolating ύie recombinant DNA molecules from the transformants and, when necessary, retrieving the inserted gene from the isolated recombinant DNA. The Notch sequences provided by the instant invention include those nucleotide sequences encoding substantially the same amino acid sequences as found in native Notch protein, and those encoded amino acid sequences with functionally equivalent amino acids, all as described in Section 5.6 infra for Notch derivatives. Similar methods to those described supra can be used to obtain a nucleic acid encoding Delta, Serrate, or adhesive portions thereof, or other toporythmic gene of interest. In a specific embodiment, the Delta nucleic acid has at least a portion of the sequence shown in Figure 1 (SEQ ID NO: l). In another specific embodiment, the Serrate nucleic acid has at least a portion of the sequence shown in Figure 5 (SEQ ID NO:3). The nucleic acid sequences encoding toporythmic proteins can be isolated from porcine, bovine, feline, avian, equine, or canine, as well as primate sources and any other species in which homologs of known toporythmic genes [including but not limited to the following genes (with the publication of sequences in parentheses): Delta (Vassin et al., 1987, EMBO J. 6, 3431-3440; Kopczynski et al., 1988, Genes Dev. 2, 1723- 1735; note corrections to the Kopczynski et al. sequence found in Figure 1 hereof (SEQ ID NO: l and SEQ ID NO:2)) and Serrate (Fleming et al. , 1990, Genes & Dev. 4, 2188-2201)] can be identified. Such sequences can be altered by substitutions, additions or deletions that provide for functionally equivalent molecules, as described supra. 5.8. RECOMBINANT PRODUCTION OF PROTEIN THERAPEUTICS The nucleic acid coding for a protein Therapeutic of the invention can be inserted into an appropriate expression vector, / ' . e. , a vector which contains the necessary elements for the transcription and translation of the inserted protein-coding sequence. The necessary transcriptional and translational signals can also be supplied by the native toporythmic gene and/or its flanking regions. A variety of host-vector systems may be utilized to express the protein-coding sequence. These include but are not limited to mammalian cell systems infected with virus (e.g. , vaccinia virus, adenovirus, etc.); insect cell systems infected with virus (e.g. , baculo virus); microorganisms such as yeast containing yeast vectors, or bacteria transformed with bacteriophage, DNA, plasmid DNA, or cosmid DNA. The expression elements of vectors vary in their strengths and specificities. Depending on the host-vector system utilized, any one of a number of suitable transcription and translation elements may be used. In a specific embodiment, the adhesive portion of the Notch gene, e.g. , that encoding EGF- like repeats (ELR) 11 and 12, is expressed. In other specific embodiments, the human Notch gene is expressed, or a sequence encoding a functionally active portion of human Notch. Any of the methods previously described for the insertion of DNA fragments into a vector may be used to construct expression vectors containing a chimeric gene consisting of appropriate transcriptional/translational control signals and the protein coding sequences. These methods may include in vitro recombinant DNA and synthetic techniques and in vivo recombinants (genetic recombination). Expression of nucleic acid sequence encoding a Notch protein or peptide fragment may be regulated by a second nucleic acid sequence so that the Notch protein or peptide is expressed in a host transformed with the recombinant DNA molecule. For example, expression of a Notch protein may be controlled by any promoter/enhancer element known in the art. Promoters which may be used to control toporythmic gene expression include, but are not limited to, the SV40 early promoter region (Bernoist and Chambon, 1981 , Nature 290, 304- 310), the promoter contained in the 3' long terminal repeat of Rous sarcoma virus (Yamamoto, et al., 1980, Cell 22, 787-797), the herpes thymidine kinase promoter (Wagner et al., 1981 , Proc. Natl. Acad. Sci. U.S.A. 78, 1441-1445), the regulatory sequences of the metallothionein gene (Brinster et al. , 1982, Nature 296, 39-42); prokaryotic expression vectors such as the β-lactamase promoter (Villa-Kamaroff, et al., 1978, Proc. Natl. Acad. Sci. U.S.A. 75, 3727-3731), or the tac promoter (DeBoer, et al., 1983, Proc. Natl. Acad. Sci. U.S.A. 80, 21- 25); see also "Useful proteins from recombinant bacteria" in Scientific American, 1980, 242, 74-94; plant expression vectors comprising the nopaline synthetase promoter region (Herrera-Estrella et al., Nature 303, 209-213) or the cauliflower mosaic virus 35S RNA promoter (Gardner, et al., 1981 , Nucl. Acids Res. 9, 2871), and the promoter of the photosynthetic enzyme ribulose biphosphate carboxylase (Herrera-Estrella et al. , 1984, Nature 310, 115-120); promoter elements from yeast or other fungi such as the Gal 4 promoter, the ADC (alcohol dehydrogenase) promoter, PGK (phosphoglycerol kinase) promoter, alkaline phosphatase promoter, and the following animal transcriptional control regions, which exhibit tissue specificity and have been utilized in transgenic animals: elastase I gene control region which is active in pancreatic acinar cells (Swift et al., 1984, Cell 38, 639-646; Ornitz et al., 1986, Cold Spring Harbor Symp. Quant. Biol. 50, 399-409; MacDonald, 1987, Hepatology 7, 425-515); insulin gene control region which is active in pancreatic beta cells (Hanahan, 1985, Nature 315, 115-122), immunoglobulin gene control region which is active in lymphoid cells (Grosschedl et al. , 1984, Cell 38, 647-658; Adames et al., 1985, Nature 318, 533-538; Alexander et al. , 1987, Mol. Cell. Biol. 7, 1436-1444). mouse mammary tumor virus control region which is active in testicular, breast. lymphoid and mast cells (Leder et al. , 1986, Cell 45, 485-495), albumin gene control region which is active in liver (Pinkert et al. , 1987. Genes and Devel. 1 , 268-276), alpha-fetoprotein gene control region which is active in liver (Krumlauf et al., 1985, Mol. Cell. Biol. 5, 1639-1648; Hammer et al., 1987, Science 235, 53-58; alpha 1-antitrypsin gene control region which is active in the liver (Kelsey et al., 1987, Genes and Devel. 1 , 161-171), beta-globin gene control region which is active in myeloid cells (Mogram et al., 1985, Nature 315, 338-340; Kollias et al., 1986, Cell 46, 89-94; myelin basic protein gene control region which is active in oligodendrocyte cells in the brain (Readhead et al. , 1987, Cell 48, 703-712); myosin light chain-2 gene control region which is active in skeletal muscle (Sani, 1985, Nature 314, 283-286). and gonadotropic releasing hormone gene control region which is active in the hypothalamus (Mason et al., 1986, Science 234, 1372-1378). Expression vectors containing Notch gene inserts can be identified by three general approaches: (a) nucleic acid hybridization, (b) presence or absence of "marker" gene functions, and (c) expression of inserted sequences. In the first approach, the presence of a foreign gene inserted in an expression vector can be detected by nucleic acid hybridization using probes comprising sequences that are homologous to an inserted toporythmic gene. In the second approach, the recombinant vector/host system can be identified and selected based upon me presence or absence of certain "marker" gene functions (e.g. , thymidine kinase activity, resistance to antibiotics, transformation phenotype, occlusion body formation in baculovirus, etc.) caused by the insertion of foreign genes in the vector. For example, if the Notch gene is inserted within die marker gene sequence of the vector, recombinants containing the Notch insert can be identified by the absence of the marker gene function. In the third approach, recombinant expression vectors can be identified by assaying the foreign gene product expressed by the recombinant. Such assays can be based, for example, on the physical or functional properties of the Notch gene product in in vitro assay systems, e.g.. aggregation (adhesive) ability (see Sections 6-7, infra). Once a particular recombinant DNA molecule is identified and isolated, several me ods known in the art may be used to propagate it. Once a suitable host system and growth conditions are established, recombinant expression vectors can be propagated and prepared in quantity. As previously explained, the expression vectors which can be used include, but are not limited to, the following vectors or their derivatives: human or animal viruses such as vaccinia virus or adenovirus: insect viruses such as baculovirus; yeast vectors; bacteriophage vectors (e.g. , lambda), and plasmid and cosmid DNA vectors, to name but a few. in addition, a host cell strain may be chosen which modulates the expression of the inserted sequences, or modifies and processes the gene product in the specific fashion desired. Expression from certain promoters can be elevated in the presence of certain inducers; thus, expression of the genetically engineered Notch protein may be controlled. Furthermore, different host cells have characteristic and specific mechanisms for the translational and post- translational processing and modification (e.g. , glycosylation, cleavage) of proteins. Appropriate cell lines or host systems can be chosen to ensure the desired modification and processing of the foreign protein expressed. For example, expression in a bacterial system can be used to produce an unglycosylated core protein product. Expression in yeast will produce a glycosylated product. Expression in mammalian cells can be used to ensure "native" glycosylation of a heterologous mammalian toporythmic protein. Furthermore, different vector/host expression systems may effect processing reactions such as proteolytic cleavages to different extents. In other specific embodiments, the Notch protein, fragment, analog, or derivative may be expressed as a fusion, or chimeric protein product (comprising the protein, fragment, analog, or derivative joined via a peptide bond to a heterologous protein sequence (of a different protein)). Such a chimeric product can be made by ligating the appropriate nucleic acid sequences encoding me desired amino acid sequences to each other by methods known in me art, in the proper coding frame, and expressing the chimeric product by methods commonly known in the art. Alternatively, such a chimeric product may be made by protein synthetic techniques, e.g. , by use of a peptide synthesizer. Both cDNA and genomic sequences can be cloned and expressed. In other embodiments, a Notch cDNA sequence may be chromosomally integrated and expressed. Homologous recombination procedures known in the art may be used. 5.8.1. IDENTIFICATION AND PURIFICATION OF THE EXPRESSED GENE PRODUCT Once a recombinant which expresses the Notch gene sequence is identified, the gene product may be analyzed. This can be achieved by assays based on the physical or functional properties of the product, including radioactive labelling of the product followed by analysis by gel electrophoresis. Once the Notch protein is identified, it may be isolated and purified by standard methods including chromatography (e.g. , ion exchange, affinity, and sizing column chromatography), centrifugation, differential solubility, or by any other standard technique for the purification of proteins. The functional properties may be evaluated using any suitable assay, including, but not limited to, aggregation assays (see Sections 6-7). 5.9. DERIVATIVES AND ANALOGS OF NOTCH AND OTHER TOPORYTHMIC PROTEINS The invention further provides, as Therapeutics, derivatives (including but not limited to fragments) and analogs of Notch proteins. Also provided as Therapeutics are other toporythmic proteins and derivatives and analogs thereof, or Notch ligands, in particular, which promote or, alternatively, inhibit the interactions of such other toporythmic proteins with Notch. The production and use of derivatives and analogs related to Notch are within the scope of the present invention. In a specific embodiment, the derivative or analog is functionally active, i.e.. capable of exhibiting one or more functional activities associated widi a full-length, wild-type Notch protein. As one example, such derivatives or analogs which have the desired antigenicity can be used, for example, in diagnostic immunoassays as described in Section 5.3. Molecules which retain, or alternatively inhibit, a desired Notch property, e.g. , binding to Delta or otiier toporythmic proteins, binding to a intracellular ligand, can be used therapeutically as inducers, or inhibitors, respectively, of such property and its physiological correlates. Derivatives or analogs of Notch can be tested for the desired activity by procedures known in the art, including but not limited to the assays described infra. In one specific embodiment, peptide libraries can be screened to select a peptide with the desired activity; such screening can be carried out by assaying, e.g. , for binding to Notch or a Notch binding partner such as Delta. In particular, Notch derivatives can be made by altering Notch sequences by substitutions, additions or deletions that provide for functionally equivalent molecules. Due to the degeneracy of nucleotide coding sequences, other DNA sequences which encode substantially the same amino acid sequence as a Notch gene may be used in the practice of the present invention. These include but are not limited to nucleotide sequences comprising all or portions of Notch genes which are altered by the substitution of different codons tiiat encode a functionally equivalent amino acid residue within the sequence, thus producing a silent change. Likewise, die Notch derivatives of the invention include, but are not limited to, those containing, as a primary amino acid sequence, all or part of the amino acid sequence of a Notch protein including altered sequences in which functionally equivalent amino acid residues are substituted for residues within the sequence resulting in a silent change. For example, one or more amino acid residues within the sequence can be substituted by another amino acid of a similar polarity which acts as a functional equivalent, resulting in a silent alteration. Substitutes for an amino acid within the sequence may be selected from other members of the class to which the amino acid belongs. For example, the nonpolar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan and methionine. The polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine. The positively charged (basic) amino acids include arginine, lysine and histidine. The negatively charged (acidic) amino acids include aspartic acid and glutamic acid. Derivatives or analogs of Notch include but are not limited to those peptides which are substantially homologous to Notch or fragments thereof, or whose encoding nucleic acid is capable of hybridizing to a Notch nucleic acid sequence. The Notch derivatives and analogs of the invention can be produced by various methods known in the art. The manipulations which result in their production can occur at the gene or protein level. For example, the cloned Notch gene sequence can be modified by any of numerous strategies known in the art (Maniatis, T. , 1989, Molecular Cloning, A Laboratory Manual, 2d ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, New York). The sequence can be cleaved at appropriate sites with restriction endonuclease(s), followed by further enzymatic modification if desired, isolated, and ligated in vitro. In the production of die gene encoding a derivative or analog of Notch, care should be taken to ensure that the modified gene remains within the same translational reading frame as Notch, uninterrupted by translational stop signals, in the gene region where the desired Notch activity is encoded. Additionally, the Notch-encoding nucleic acid sequence can be mutated in vitro or in vivo, to create and/or destroy translation, initiation, and/or termination sequences, or to create variations in coding regions and/or form new restriction endonuclease sites or destroy preexisting ones, to facilitate further in vitro modification. Any technique for mutagenesis known in the art can be used, including but not limited to, in vitro site-directed mutagenesis (Hutchinson, C, et al., 1978, J. Biol. Chem 253:6551), use of TAB ® linkers (Pharmacia), etc. Manipulations of the Notch sequence may also be made at the protein level. Included within the scope of the invention are Notch protein fragments or other derivatives or analogs which are differentially modified during or after translation, e.g. , by glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to an antibody molecule or other cellular ligand, etc. Any of numerous chemical modifications may be carried out by known techniques, including but not limited to specific chemical cleavage by cyanogen bromide, trypsin, chymotrypsin, papain, V8 protease, NaBH 4 ; acetylation, formylation. oxidation, reduction; metabolic synthesis in the presence of tunicamycin; etc. In addition, analogs and derivatives of Notch can be chemically synthesized. For example, a peptide corresponding to a portion of a Notch protein which comprises the desired domain, or which mediates the desired aggregation activity in vitro, or binding to a receptor, can be synthesized by use of a peptide synthesizer. Furthermore, if desired, nonclassical amino acids or chemical amino acid analogs can be introduced as a substitution or addition into the Notch sequence. Non-classical amino acids include but are not limited to the D-isomers of the common amino acids, α-amino isobutyric acid, 4-aminobutyric acid, hydroxyproline, sarcosine, citrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, 0-alanine, designer amino acids such as 3-methyl amino acids, Cα-methyl amino acids, and Nα-methyl amino acids. In a specific embodiment, the Notch derivative is a chimeric, or fusion, protein comprising a Notch protein or fragment thereof fused via a peptide bond at its amino- and/or carboxy-terminus to a non-Notch amino acid sequence. In one embodiment, such a chimeric protein is produced by recombinant expression of a nucleic acid encoding me protein (comprising a Notch-coding sequence joined in-frame to a non-Notch coding sequence). Such a chimeric product can be made by ligating the appropriate nucleic acid sequences encoding the desired amino acid sequences to each other by methods known in the art, in the proper coding frame, and expressing the chimeric product by methods commonly known in the art. Alternatively, such a chimeric product may be made by protein synthetic techniques, e.g. , by use of a peptide synthesizer. In a specific embodiment, a chimeric nucleic acid encoding a mature Notch protein with a heterologous signal sequence is expressed such that the chimeric protein is expressed and processed by the cell to the mature Notch protein. As another example, and not by way of limitation, a recombinant molecule can be constructed according to the invention, comprising coding portions of bo Notch and another toporythmic gene, e.g. , Delta. The encoded protein of such a recombinant molecule could exhibit properties associated with both Notch and Delta and portray a novel profile of biological activities, including agonists as well as antagonists. The primary sequence of Notch and Delta may also be used to predict tertiary structure of the molecules using computer simulation (Hopp and Woods, 1981 , Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828): Notch/Delta chimeric recombinant genes could be designed in light of correlations between tertiary structure and biological function. Likewise, chimeric genes comprising portions of Notch fused to any heterologous (non-Notch) protein-encoding sequences may be constructed. A specific embodiment relates to a chimeric protein comprising a fragment of Notch of at least six amino acids. In anotfier specific embodiment, the Notch derivative is a fragment of Notch comprising a region of homology with another toporythmic protein. As used herein, a region of a first protein shall be considered "homologous" to a second protein when the amino acid sequence of the region is at least 30% identical or at least 75% either identical or involving conservative changes, when compared to any sequence in the second protein of an equal number of amino acids as the number contained in the region. Derivatives of Serrate, Delta, other toporythmic proteins, and the adhesive portions thereof, can be made by methods similar to those described supra. 5.9.1. DERIVATIVES OF NOTCH CONTAINING ONE OR MORE DOMAINS OF THE PROTEIN In a specific embodiment, the invention provides Therapeutics that are Notch derivatives and analogs, in particular Notch fragments and derivatives of such fragments, mat comprise one or more domains of the Notch protein, including but not limited to the extracellular domain, transmembrane domain, intracellular domain, membrane-associated region, one or more of the EGF-like repeats (ELR) of the Notch protein, the cdclO repeats, and the Notch/lin-12 repeats. In specific embodiments, the Notch derivative may lack all or a portion of the ELRs, or one or more other regions of the protein. In a specific embodiment, relating to a Notch protein of a species other than D. melanogaster, preferably human, fragments comprising specific portions of Notch are those comprising portions in the respective Notch protein most homologous to specific fragments of the Drosophila Notch protein (e.g. , ELR 11 and ELR 12). 5.9.2. DERIVATIVES OF NOTCH OR OTHER TOPORYTHMIC PROTEINS THAT MEDIATE BINDING TO TOPORYTHMIC PROTEIN DOMAINS. AND INHIBITORS THEREOF The invention also provides Notch fragments, and analogs or derivatives of such fragments, which mediate binding to toporythmic proteins (and d us are termed herein "adhesive"), and nucleic acid sequences encoding the foregoing. Also included as Therapeutics of the invention are toporythmic (e.g. , Delta, Serrate) protein fragments, and analogs or derivatives thereof, which mediate heterotypic binding to Notch (and thus are termed herein "adhesive"), and nucleic acid sequences relating to the foregoing. Also included as Therapeutics of the invention are inhibitors (e.g. , peptide inhibitors) of the foregoing toporythmic protein interactions with Notch. The ability to bind to a toporythmic protein can be demonstrated by in vitro aggregation assays with cells expressing such a toporythmic protein as well as cells expressing Notch or a Notch derivative (See Section 6). That is, the ability of a protein fragment to bind to a Notch protein can be demonstrated by detecting the ability of the fragment, when expressed on the surface of a first cell, to bind to a Notch protein expressed on the surface of a second cell. Inhibitors of the foregoing interactions can be detected by their ability to inhibit such aggregation in vitro. The nucleic acid sequences encoding toporythmic proteins or adhesive domains thereof, for use in such assays, can be isolated from human, porcine, bovine, feline, avian, equine, canine, or insect, as well as primate sources and any other species in which homologs of known toporythmic genes can be identified. In a specific embodiment, the adhesive fragment of Notch is that comprising the portion of Notch most homologous to ELR 11 and 12, i.e., amino acid numbers 447 through 527 (SEQ ID NO: 14) of the Drosophila Notch sequence (see Figure 4). In yet another specific embodiment, the adhesive fragment of Delta mediating binding to Notch is that comprising the portion of Delta most homologous to about amino acid numbers 1-230 of the Drosophila Delta sequence (SEQ ID NO:2). In a specific embodiment relating to an adhesive fragment of Serrate, such fragment is that comprising the portion of Serrate most homologous to about amino acid numbers 85-283 or 79-282 of the Drosophila Serrate sequence (see Figure 5 (SEQ ID NO:4)). Due to the degeneracy of nucleotide coding sequences, other DNA sequences which encode substantially die same amino acid sequence as the adhesive sequences may be used in the practice of the present invention. These include but are not limited to nucleotide sequences comprising all or portions of the Notch. Delta, or Serrate genes which are altered by the substitution of different codons that encode a functionally equivalent amino acid residue within the sequence, thus producing a silent change. Likewise, the adhesive protein fragments or derivatives thereof, of the invention include, but are not limited to, those containing, as a primary amino acid sequence, all or part of the amino acid sequence of the adhesive domains including altered sequences in which functionally equivalent amino acid residues are substituted for residues within the sequence resulting in a silent change. Adhesive fragments of toporythmic proteins and potential derivatives, analogs or peptides related to adhesive toporythmic protein sequences, can be tested for the desired binding activity e.g. , by the in vitro aggregation assays described in the examples herein. Adhesive derivatives or adhesive analogs of adhesive fragments of toporythmic proteins include but are not limited to those peptides which are substantially homologous to the adhesive fragments, or whose encoding nucleic acid is capable of hybridizing to the nucleic acid sequence encoding the adhesive fragments, and which peptides and peptide analogs have positive binding activity e.g. , as tested in vitro by an aggregation assay such as described in die examples sections infra. Such derivatives and analogs are envisioned as Therapeutics and are within the scope of the present invention. The adhesive-protein related derivatives, analogs, and peptides of die invention can be produced by various mediods known in the art. The manipulations which result in their production can occur at d e gene or protein level (see Section 5.6). Additionally, the adhesive-encoding nucleic acid sequence can be mutated in vitro or in vivo; and manipulations of the adhesive sequence may also be made at the protein level (see Section 5.6). In addition, analogs and peptides related to adhesive fragments can be chemically synthesized. 5.10. ASSAYS OF NOTCH PROTEINS, 10 DERIVATIVES AND ANALOGS The in vitro activity of Notch proteins, derivatives and analogs, and other toporythmic proteins which bind to Notch, can be assayed by various methods. For example, in one embodiment, where one is assaying for the J5 ability to bind or compete wi wild-type Notch for binding to anti-Notch antibody, various immunoassays known in the art can be used, including but not limited to competitive and non-competitive assay systems using techniques such as radioimmunoassays, ELISA (enzyme linked immunosorbent assay), "sandwich" immunoassays, immunoradiometric assays, gel diffusion precipitin reactions, 0 immunodiffusion assays, ]n sjtu immunoassays (using colloidal gold, enzyme or radioisotope labels, for example), western blots, precipitation reactions, agglutination assays (e.g. , gel agglutination assays, hemagglutination assays), complement fixation assays, immunofluorescence assays, protein A assays, and immunoelectrophoresis assays, etc. In one embodiment, antibody binding is detected by detecting a label on me primary antibody. In another embodiment, the primary antibody is detected by detecting binding of a secondary antibody or reagent to the primary antibody. In a further embodiment, the secondary antibody is labelled. Many means are known in the art for detecting binding in an immunoassay and are within the scope of the present invention. 0 In another embodiment, where one is assaying for ie ability to mediate binding to Notch, one can carry out an in vitro aggregation assay such as 35 described infra in Section 6 or 7 (see also Fehon et al., 1990, Cell 61:523-534; Rebay et al., 1991, Cell 67:687-699). In another embodiment, where another ligand for Notch is identified, ligand binding can be assayed, e.g. , by binding assays well known in 5 the art. In another embodiment, physiological correlates of ligand binding to cells expressing a Notch receptor (signal transduction) can be assayed. In another embodiment, in insect or other model systems, genetic studies can be done to study die phenotypic effect of a Notch mutant that is a derivative or analog of wild-type Notch. 10 Odier methods will be known to the skilled artisan and are within the scope of die invention. 5.11. ANTIBODIES TO NOTCH PROTEINS, DERIVATIVES AND ANALOGS J5 According to one embodiment of the invention, antibodies and fragments containing the binding domain thereof, directed against Notch are Therapeutics. Accordingly, Notch proteins, fragments or analogs or derivatives mereof, in particular, human Notch proteins or fragments iereof, may be used as immunogens to generate anti-Notch protein antibodies. Such antibodies can be 0 polyclonal, monoclonal, chimeric, single chain, Fab fragments, or from an Fab expression library. In a specific embodiment, antibodies specific to EGF-like repeats 11 and 12 of Notch may be prepared. In other embodiments, antibodies reactive with the extracellular domain of Notch can be generated. One example of such antibodies may prevent aggregation in an in vitro assay. In another 5 embodiment, antibodies specific to human Notch are produced. Various procedures known in ie art may be used for the production of polyclonal antibodies to a Notch protein or peptide. In a particular embodiment, rabbit polyclonal antibodies to an epitope of the human Notch protein encoded by a sequence depicted in Figure 10 or 11, or a subsequence 0 thereof, can be obtained. For the production of antibody, various host animals can be immunized by injection with the native Notch protein, or a syn etic version, or fragment thereof, including but not limited to rabbits, mice, rats, etc. 5 Various adjuvants may be used to increase the immunological response, depending on the host species, and including but not limited to Freund's (complete and incomplete), mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhold limpet hemocyanins, dinitrophenol, and potentially useful human adjuvants such as BCG (bacille Calmette-Guerin) and corynebacterium parvum. For preparation of monoclonal antibodies directed toward a Notch protein sequence, any technique which provides for the production of antibody molecules by continuous cell lines in culture may be used. For example, the hybridoma technique originally developed by Kohler and Milstein (1975, Nature 256, 495-497), as well as the trioma technique, the human B-cell hybridoma technique (Kozbor et al., 1983, Immunology Today 4, 72), and the EBV- hybridoma technique to produce human monoclonal antibodies (Cole et al., 1985, in Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96). Antibody fragments which contain die idiotype (binding domain) of the molecule can be generated by known techniques. For example, such fragments include but are not limited to: the F(ab') 2 fragment which can be produced by pepsin digestion of die antibody molecule; the Fab' fragments which can be generated by reducing the disulfide bridges of the F(ab') 2 fragment, and die Fab fragments which can be generated by treating the antibody molecule with papain and a reducing agent. In die production of antibodies, screening for die desired antibody can be accomplished by techniques known in the art, e.g. ELISA (enzyme-linked immunosorbent assay). For example, to select antibodies which recognize the adhesive domain of a Notch protein, one may assay generated hybridomas for a product which binds to a protein fragment containing such domain. For selection of an antibody specific to human Notch, one can select on the basis of positive binding to human Notch and a lack of binding to Drosophila Notch. in addition to therapeutic utility, the foregoing antibodies have utility in diagnostic immunoassays as described in Section 5.6 supra. Similar procedures to those described supra can be used to make Therapeutics which are antibodies to domains of otiier proteins (particularly toporythmic proteins) that bind or otherwise interact with Notch (e.g. , adhesive fragments of Delta or Serrate). 6. DOMAINS OF NOTCH MEDIATE BINDING WITH DELTA Intermolecular association between the products of the Notch and Delta genes was detected by studying the effects of their expression on aggregation in Drosophila Schneider's 2 (S2) cells (Fehon et al., 1990, Cell 61, 523-534). Direct evidence of intermolecular interactions between Notch and Delta is described herein, as well as an assay system that can be used in dissecting die components of this interaction. Normally nonadhesive Drosophila S2 cultured cells that express Notch bind specifically in a calcium-dependent manner to cells that express Delta. Furthermore, while cells that express Notch do not bind to one another, cells that express Delta do bind to one another, suggesting that Notch and Delta can compete for binding to Delta at the cell surface. Notch and Delta form detergent-soluble complexes both in cultured cells and embryonic cells, suggesting that Notch and Delta interact directly at die molecular level in vitro and in vivo. The analyses suggest that Notch and Delta proteins interact at die cell surface via their extracellular domains. 6.1. EXPERIMENTAL PROCEDURES 6.1.1. EXPRESSION CONSTRUCTS Expression constructs are described in Fehon et al., 1990, Cell 61:523-534. Briefly, Notch encoded by the Mglla minigene a cDNA/genomic chimeric construct (Ramos et al., 1989. Genetics 123, 337-348) was expressed following insertion into pRmHa-3 (Bunch, et al., 1988, Nucl. Acids Res. 16, 1043-1061). In the resulting construct, designated pMtNMg, the metallothionein promoter in pRmHa-3 is fused to Notch sequences starting 20 nucleotides upstream of the translation start site. The extracellular Notch construct (ECN1), was derived from a Notch cosmid (Ramos et al., 1989, Genetics 123, 337-348), and has an internal deletion of the Notch coding sequences from amino acids 1790 to 2625 inclusive (Wharton et al., 1985, Cell 43, 567-581), and a predicted frameshift that produces a novel 59 amino acid carboxyl terminus. For the Delta expression construct, the Dll cDNA (Kopczynski et al., 1988, Genes Dev. 2, 1723-1735; Figure 1; SEQ ID NO: l), which includes die complete coding capacity for Delta, was inserted into die EcoRI site of pRmHa-3. This construct was called pMTDll. 6.1.2. ANTIBODY PREPARATION Hybridoma cell line C17.9C6 was obtained from a mouse immunized with a fusion protein based on a 2.1 kb Sall-Hindlll fragment that includes coding sequences for most of the intracellular domain of Notch (amino acids 1791-2504; Wharton et al., 1985, Cell 43, 567-581). The fragment was subcloned into pUR289 (Ruther and Muller-Hill, 1983, EMBO J. 2, 1791-1794), and dien transferred into die pATH 1 expression vector (Dieckmann and Tzagoloff, 1985, J. Biol. Chem. 260, 1513-1520) as a Bglll-Hindlll fragment. Soluble fusion protein was expressed, precipitated by 25% (NH 4 ) 2 SO 4 , resuspended in 6 M urea, and purified by preparative isoelectric focusing using a Rotofor (Bio-Rad) (for details, see Fehon, 1989, Rotofor Review No. 7, Bulletin 1518, Richmond, California: Bio-Rad Laboratories). Mouse polyclonal antisera were raised against the extracellular domain of Notch using four BstYl fragments of 0.8 kb (amino acids 237-501 : Wharton et al., 1985, Cell 43, 567-581), 1.1 kb (amino acids 501-868), 0.99 kb (amino acids 868-1200), and 1.4 kb (amino acids 1465-1935) length, which spanned from the fifth EGF-like repeat across the transmembrane domain, singly inserted in-frame into the appropriate pGEX expression vector (Smith and Johnson, 1988, Gene 67, 31-40). Fusion proteins were purified on glutathione- agarose beads (SIGMA). Mouse and rat antisera were precipitated with 50% (NH 4 ) 2 SO 4 and resuspended in PBS (150 mM NaCl, 14 mM Na 2 HPO 4 , 6 mM NaH 2 PO 4 ) with 0.02% NaN 3 . Hybridoma cell line 201 was obtained from a mouse immunized widi a fusion protein d at includes coding sequences from the extracellular domain of Delta (Kopczynski et al., 1988, Genes Dev. 2, 1723-1735), including sequences extending from the fourth through the ninth EGF-like repeats in Delta (amino acids 350-529). Rat polyclonal antisera were obtained following immunization with antigen derived from the same fusion protein construct. In this case, fusion protein was prepared by lysis of IPTG-induced cells in SDS-Laemmli buffer (Carroll and Laughon, 1987, in DNA Cloning, Volume III, D.M. Glover, ed. (Oxford: IRL Press), pp. 89-111), separation of proteins by SDS-PAGE, excision of die appropriate band from die gel, and electroelution of antigen from the gel slice for use in immunization (Harlow and Lane, 1988, Antibodies: A Laboratory Manual (Cold Spring Harbor, New York: Cold Spring Harbor Laboratory)). 6.1.3. CELL CULTURE AND TRANSFECTION The S2 cell line (Schneider, 1972, J. Embryol. Exp. Moφh. 27, 353-365) was grown in M3 medium (prepared by Hazleton Co.) supplemented with 2.5 mg/ml Bacto-Peptone (Difco), 1 mg/ml TC Yeastolate (Difco), 11 % heat- inactivated fetal calf serum (FCS) (Hyclone), and 100 U/ml penicillin-100 μg/ml streptomycin-0.25 μg/ml fungizone (Hazleton). Cells growing in log phase at ~ 2 x IO 6 cells/ml were transfected with 20 μg of DNA-calcium phosphate coprecipitate in 1 ml per 5 ml of culture as previously described (Wigler et al., 1979, Proc. Natl. Acad. Sci. USA 78, 1373-1376), with the exception that BES buffer (SIGMA) was used in place of HEPES buffer (Chen and Okayama, 1987, Mol. Cell. Biol. 7, 2745-2752). After 16-18 hr, cells were transferred to conical centrifuge tubes, pelleted in a clinical centrifuge at full speed for 30 seconds, rinsed once with 1/4 volume of fresh complete medium, resuspended in their original volume of complete medium, and returned to the original flask. Transfected cells were dien allowed to recover for 24 hr before induction. 6.1.4. AGGREGATION ASSAYS Expression of the Notch and Delta metal lodiionein constructs was induced by the addition of CuSO 4 to 0.7 mM. Cells transfected with the ECN1 construct were treated similarly. Cells were then mixed, incubated under aggregation conditions, and scored for dieir ability to aggregate using specific antisera and immuno fluorescence microscopy to visualize expressing cells. Two types of aggregation assays were used. In the first assay, a total of 3 ml of cells (5-10 x IO 6 cells/ml) was placed in a 25 ml Erlenmeyer flask and rotated at 40-50 φm on a rotary shaker for 24-48 hr at room temperature. For these experiments, cells were mixed 1-4 hr after induction began and induction was continued throughout the aggregation period. In the second assay, — 0.6 ml of cells were placed in a 0.6 ml Eppendorf tube (leaving a small bubble) after an overnight induction (12-16 hr) at room temperature and rocked gently for 1-2 hr at 4°C. The antibody inhibition and Ca 2+ dependence experiments were performed using the latter assay. For Ca 2+ dependence experiments, cells were first collected and rinsed in balanced saline solution (BSS) with 11 % FCS (BSS- FCS; FCS was dialyzed against 0.9% NaCl, 5mM Tris [pH 7.5]) or in Ca 2+ free BSS-FCS containing 10 mM EGTA (Snow et al. , 1989, Cell 59, 313-323) and then resuspended in the same medium at the original volume. For the antibody inhibition experiments, Notch-transfected cells were collected and rinsed in M3 medium and then treated before aggregation in M3 medium for 1 hr at 4°C with a 1:250 dilution of immune or preimmune sera from each of die four mice immunized with fusion proteins containing segments from the extracellular domain of Notch (see Antibody Preparation above). 6.1.5. IMMUNOFLUORESCENCE Cells were collected by centrifugation (3000 φm for 20 seconds in an Eppendorf microcentrifuge) and fixed in 0.6 ml Eppendorf tubes widi 0.5 ml of freshly made 2% paraformaldehyde in PBS for 10 min at room temperature. After fixing, cells were collected by centrifugation, rinsed twice in PBS, and stained for 1 hr in primary antibody in PBS with 0.1 % saponin (SIGMA) and 1 % normal goat serum (Pocono Rabbit Farm. Canadensis. PA). Monoclonal antibody supernatants were diluted 1: 10 and mouse or rat sera were diluted 1 : 1000 for this step. Cells were then rinsed once in PBS and stained for 1 hr in specific secondary antibodies (double-labeling grade goat anti-mouse and goat anti-rat, Jackson Immunoresearch) in PBS-saponin-normal goat serum. After this incubation, cells were rinsed twice in PBS and mounted on slides in 90% glycerol, 10% 1 M Tris (pH 8.0), and 0.5% n-propyl gallate. Cells were viewed under epifluorescence on a Leitz Orthoplan 2 microscope. Confocal micrographs were taken using the Bio-Rad MRC 500 system connected to a Zeiss Axiovert compound microscope. Images were collected using the BHS and GHS filter sets, aligned using the ALIGN program, and merged using MERGE. Fluorescent bleed-through from the green into the red channel was reduced using the BLEED program (all software provided by Bio-Rad). Photographs were obtained directly from the computer monitor using Kodak Ektar 125 film. 6.1.6. CELL LYSATES, IMMUNOPRECIPITATIONS, AND WESTERN BLOTS Nondenaturing detergent lysates of tissue culture and wild-type Canton-S embryos were prepared on ice in — 10 cell vol of lysis buffer (300 mM NaCl, 50 mM Tris [pH 8.0], 0.5 % NP-40, 0.5% deoxycholate, 1 mM CaCl 2 , 1 mM MgCl 2 ) with 1 mM phenylmethysulfonyl fluoride (PMSF) and diisopropyl fluorophosphate diluted 1:2500 as protease inhibitors. Lysates were sequentially triturated using 18G, 21G, and 25G needles attached to 1 cc tuberculin syringes and dien centrifuged at full speed in a microfuge 10 min at 4°C to remove insoluble material. Immunoprecipitation was performed by adding — 1 μg of antibody (1-2 μl of polyclonal antiserum) to 250-500 μl of cell lysate and incubating for 1 hr at 4°C with agitation. To this mixture, 15 μg of goat anti- mouse antibodies (Jackson Immunoresearch; these antibodies recognize both mouse and rat IgG) were added and allowed to incubate for 1 hr at 4°C with agitation. This was followed by die addition of 100 μl of fixed Staphylococcus aureus (Staph A) bacteria (Zysorbin, Zymed; resuspended according to manufacturer's instructions), which had been collected, washed five times in lysis buffer, and incubated for another hour. Staph A-antibody complexes were then pelleted by centrifugation and washed diree times in lysis buffer followed by two 15 min washes in lysis buffer. After being transferred to a new tube, precipitated material was suspended in 50 μl of SDS-PAGE sample buffer, boiled immediately for 10 min, run on 3%-15% gradient gels, blotted to nitrocellulose, and detected using monoclonal antibodies and HRP-conjugated goat anti-mouse secondary antibodies as previously described (Johansen et al. , 1989, J. Cell Biol. 109, 2427- 2440). For total cellular protein samples used on Western blots, cells were 0 collected by centrifugation, lysed in 10 cell vol of sample buffer that contained 1 mM PMSF, and boiled immediately. 6.2. RESULTS 6.2.1. THE EXPRESSION OF NOTCH AND 5 DELTA IN CULTURED CELLS To detect interactions between Notch and Delta, we examined the behavior of cells expressing these proteins on their surfaces using an aggregation assay. We chose the S2 cell line (Schneider, 1972, J. Embryol. Exp. Moφh. 27, 353-365) for these studies. S2 cells express an aberrant Notch message and no o detectable Notch due to a rearrangement of the 5' end of the Notch coding sequence. These cells also express no detectable Delta. Results of Western blot and immunofluorescent analysis clearly showed diat die Notch and Delta constructs support expression of proteins of the expected sizes and subcellular localization. 5 6.2.2. CELLS THAT EXPRESS NOTCH AND DELTA AGGREGATE A simple aggregation assay was used to detect interactions between Notch and Delta expressed on the surface of S2 cells. S2 cells in log phase growth were separately transfected with 0 eidier the Notch or Delta metallothionein promoter construct. After induction with CuSO 4 , transfected cells were mixed in equal numbers and allowed to aggregate overnight at room temperature (for details, see Experimental 5 Procedures, Section 6.1). Alternatively, in some experiments intended to reduce metabolic activity, cells were mixed gently at 4°C for 1-2 hr. To determine whether aggregates had formed, cells were processed for immunofluorescence microscopy using antibodies specific for each gene product and differently labeled fluorescent secondary antibodies. Expressing cells usually constituted less than 5 % of the total cell population because transient rather than stable transformants were used. The remaining cells either did not express a given protein or expressed at levels too low for detection by immunofluorescence microscopy. As controls, we performed aggregations with only a single type of transfected cell. The results (Fehon et al., 1990, Cell 61 :523-534) showed that while Notch-expressing (Notch + ) cells alone did not form aggregates in the assay, Delta-expressing (Delta "1" ) cells did. The tendency for Delta + cells to aggregate was apparent even in nonaggregated control samples, where cell clusters of 4-8 cells that probably arose from adherence between mitotic sister cells commonly occurred. However, clusters were more common after incubation under aggregation conditions (e.g. , 19% of Delta + cells in aggregates before incubation vs. 37% of Delta "1" cells in aggregates after incubation), indicating that Delta + cells are able to form stable contacts with one another in this assay. In remarkable contrast to control experiments with Notch "1" cells alone, aggregation of mixtures of Notch + and Delta "1" cells resulted in die formation of clusters of up to 20 or more cells. The fraction of expressing cells found in clusters of four or more stained cells after 24 hr of aggregation ranged from 32%-54% in mixtures of Notch + and Delta "1" cells. This range was similar to that seen for Delta " * " cells alone (37%-40%) but very different from that for Notch "1" cells alone (only 0%-5%). Although a few clusters that consisted only of Delta "1" cells were found. Notch "1" cells were never found in clusters of greater dian four to five cells unless Delta "1" cells were also present. Again, all cells within these clusters expressed either Notch or Delta, even though transfected cells composed only a small fraction of the total cell population. At 48 hr, the degree of aggregation appeared higher (63%-71 %), suggesting that aggregation had not yet reached a maximum after 24 hr under these conditions. Also, cells cotransfected widi Notch and Delta constructs (so that all transfected cells express bodi proteins) aggregated in a similar fashion under the same experimental conditions. Notch involvement in the aggregation process was directly tested by examining the effect of a mixture of polyclonal antisera directed against fusion proteins diat spanned almost the entire extracellular domain of Notch on aggregation (see Experimental Procedures, Section 6.1). To minimize artifacts mat might arise due to a metabolic response to patching of surface antigens, antibody treatment and the aggregation assay were performed at 4° C in these experiments. Notch " * " cells were incubated with either preimmune or immune mouse sera for 1 hr, Delta " * " cells were added, and aggregation was performed for 1-2 hr. While Notch "1" cells pretreated with preimmune sera aggregated widi Delta " * " cells (in one of three experiments, 23% of the Notch "1" cells were in Notch + -Delta + cell aggregates), those treated widi immune sera did not (only 2% of Notch " * " cells were in aggregates). This result suggested diat the extracellular domain of Notch was required for Notch + -Delta + cell aggregation. 6.2.3. NOTCH-DELTA-MEDIATED AGGREGATION IS CALCIUM DEPENDENT The ability of expressing cells to aggregate in the presence or absence of Ca 2+ ions was tested to determine whether there is a Ca 2+ ion requirement for Notch-Delta aggregation. To minimize possible nonspecific effects due to metabolic responses to the removal of Ca 2+ , these experiments were performed at 4°C. The results clearly demonstrated a dependence of Notch- Delta- mediated aggregation on exogenous Ca , 2 2 + 6.2.4. NOTCH AND DELTA INTERACT WITHIN A SINGLE CELL The question whether Notch and Delta are associated widiin the membrane of one cell that expresses both proteins was posed by examining the distributions of Notch and Delta in cotransfected cells. To test whether the observed colocalization was coincidental or represented a stable interaction between Notch and Delta, live cells were treated with an excess of polyclonal anti-Notch antiserum. This treatment resulted in "patching" of Notch on the surface of expressing cells into discrete patches as detected by immunofluorescence. There was a distinct correlation between the distributions 5 of Notch and Delta on the surfaces of these cells after this treatment, indicating that these proteins are associated within the membrane. 6.2.5. INTERACTIONS WITH DELTA DO NOT REQUIRE THE INTRACELLULAR DOMAIN OF NOTCH JO In addition to a large extracellular domain that contains EGF-like repeats, Notch has a sizeable intracellular (IC) domain of —940 amino acids. The IC domain includes a phosphorylation site (Kidd et al. , 1989, Genes Dev. 3, 1113-1129), a putative nucleotide binding domain, a polyglutamine stretch (Wharton et al., 1985, Cell 43, 567-581; Kidd, et al., 1986, Mol. Cell. Biol. 6, J5 3094-3108), and sequences homologous to the yeast cdclO gene, which is involved in cell cycle control in yeast (Breeden and Nasmyth, 1987, Nature 329, 651-654). A variant Notch construct was used from which coding sequences for — 835 amino acids of the IC domain, including all of the structural features noted above, had been deleted (leaving 25 membrane-proximal amino acids and a novel o 59 amino acid carboxyl terminus; see Experimental Procedures). In aggregation assays, cells that expressed the ECNl construct consistently formed aggregates with Delta " * " cells, but not with themselves, just as was observed for cells diat expressed intact Notch. Shaφ bands of ECNl staining were observed within regions of contact widi Delta " * " cells, again 5 indicating a localization of ECNl within regions of contact between cells. To test for interactions within die membrane, surface antigen co-patching experiments were conducted using cells cotransfected with the ECNl and Delta constructs. As observed for intact Notch, when ECNl was patched using polyclonal antisera against the extracellular domain of Notch, ECNl and Delta colocalized at the cell 0 surface. These results demonstrate that the observed interactions between Notch and Delta within the membrane do not require the deleted portion of the IC domain of Notch and are therefore probably mediated by the extracellular domain. 5 6.2.6. NOTCH AND DELTA FORM DETERGENT-SOLUBLE INTERMOLECULAR COMPLEXES The preceding results indicated molecular interactions between Notch and Delta present widiin the same membrane and between these proteins 5 expressed on different cells. A further test for such interactions is whether these proteins would coprecipitate from nondenaturing detergent extracts of cells that express Notch and Delta. If Notch and Delta form a stable intermolecular complex either between or within cells, then it should be possible to precipitate both proteins from cell extracts using specific antisera directed against one of O these proteins. This analysis was performed by immunoprecipitating Delta with polyclonal antisera from NP-40/deoxycholate lysates (see Experimental Procedures) of cells cotransfected with the Notch and Delta constructs that had been allowed to aggregate overnight or of 0-24 hr wild-type embryos. Coprecipitation of Notch was detected in Delta immunoprecipitates J5 from cotransfected cells and embryos. However, coprecipitating Notch appeared to be present in much smaller quantities than Delta and was therefore difficult to detect. The fact that immunoprecipitation of Delta results in the coprecipitation of Notch constitutes direct evidence diat diese two proteins form stable intermolecular completes in transfected S2 cells and in embryonic cells. 0 6.3. DISCUSSION Use of an in vitro aggregation assay that employs normally nonadhesive S2 cells showed at cells that express Notch and Delta adhere specifically to one anodier. 5 7. EGF REPEATS 11 AND 12 OF NOTCH ARE REQUIRED AND SUFFICIENT FOR NOTCH-DELTA-MEDIATED AGGREGATION The same aggregation assay was used as described in Section 6, togedier with deletion mutants of Notch to identify regions within the extracellular 30 domain of Notch necessary for interactions widi Delta. The evidence shows diat the EGF repeats of Notch are directly involved in this interaction and that only 35 two (ELR 11 and 12) of the 36 EGF repeats appear necessary. These two EGF repeats are sufficient for binding to Delta and diat the calcium dependence of Notch-Delta mediated aggregation also associates with these two repeats. Finally, the two corresponding EGF repeats from the Xenopus homolog of Notch also mediate aggregation with Delta, implying that not only has the structure of Notch been evolutionarily conserved, but also its function. These results suggest that the extracellular domain of Notch is suφrisingly modular, and could potentially bind a variety of proteins in addition to Delta. (See Rebay et al., 1991, Cell 67:687-699.) 7.1. EXPERIMENTAL PROCEDURES 7.1.1. EXPRESSION CONSTRUCTS The constructs described are all derivatives of the full length Notch expression construct #1 pMtNMg (see Section 6, supra), and were made as described (Rebay et al., 1991, Cell 67:687-699). 7.1.2. CELL CULTURE AND TRANSFECTION The Drosophila S2 cell line was grown and transfected as described in Section 6, supra. The Delta-expressing stably transformed S2 cell line L-49-6-7 (kindly established by L. Cherbas) was grown in M3 medium (prepared by Hazleton Co.) supplemented with 11 % heat inactivated fetal calf serum (FCS) (Hyclone), 100 U/ml penicillin-100 μg/ml streptomycin-0.25 μg/ml fungizone (Hazleton), 2 x 10 "7 M methotrexate, 0.1 mM hypoxanthine, and 0.016 mM thymidine. 7.1.3. AGGREGATION ASSAYS AND IMMUNOFLUORESCENCE Aggregation assays and Ca ++ dependence experiments were as described supra, Section 6. Cells were stained with the anti-Notch monoclonal antibody 9C6.C17 and anti-Delta rat polyclonal antisera (details described in Section 6, supra). Surface expression of Notch constructs in unpermeabilized cells was assayed using rat polyclonal antisera raised against the 0.8 kb (amino acids 237-501 ; Wharton et al., 1985, Cell 43, 567-581) BstYI fragment from the extracellular domain of Notch. Cells were viewed under epifluorescence on a Leitz Orthoplan 2 microscope. 7.2. RESULTS 7.2.1. EGF REPEATS 11 AND 12 OF NOTCH ARE REQUIRED FOR NOTCH-DELTA MEDIATED AGGREGATION An extensive deletion analysis was undertaken of the extracellular domain of die Notch protein, which was shown (supra, Section 6; Fehon et al., 1990, Cell 61:523-534) to be involved in Notch-Delta interactions, to identify the precise domain of Notch mediating these interactions. The ability of cells transfected with die various deletion constructs to interact with Delta was tested using the aggregation assay described in Section 6. Briefly, Notch deletion constructs were transiently transfected into Drosophila S2 cells, induced with CuSO 4 , and d en aggregated overnight at room temperature with a small amount of cells from the stably transformed Delta expressing cell line L49-6-7(Cherbas), yielding a population typically composed of — 1 % Notch expressing cells and — 5% Delta expressing cells, with the remaining cells expressing neither protein. Schematic drawings of the constructs tested and results of die aggregation experiments are shown in Figure 2. To assay the degree of aggregation, cells were stained widi antisera specific to each gene product and examined widi immunofluorescent microscopy. Aggregates were defined as clusters of four or more cells containing both Notch and Delta expressing cells, and the values shown in Figure 2 represent the percentage of all Notch expressing cells found in such clusters. All numbers reflect the average result from at least two separate transfection experiments in which at least 100 Notch expressing cell units (either single cells or clusters) were scored. The initial constructs (#2 DSph and #3 ΔCla) deleted large portions of the EGF repeats. Their inability to promote Notch-Delta aggregation suggested that the EGF repeats of Notch were involved in the interaction with Delta. A series of six in-frame Clal restriction sites was used to further dissect the region between EGF repeats 7 and 30. Due to sequence homology between repeats, five of die Clal sites occur in die same relative place wi in the EGF repeat, just after the third cysteine, while the sixth site occurs just before the first cysteine of EGF repeat 31 (Figure 3). Thus, by performing a partial Clal digestion and then religating, deletions were obtained that not only preserved die open reading frame of die Notch protein but in addition frequently maintained the structural integrity and conserved spacing, at least theoretically, of the three disulfide bonds in the chimeric EGF repeats produced by the religation (Figure 2, constructs #4-14). Unfortunately, the most 3' Clal site was resistant to digestion while die next most 3' Clal site broke between EGF repeats 30 and 31. Therefore, when various Clal digestion fragments were reinserted into the framework of die complete Clal digest (construct #3 ΔCla), die overall structure of the EGF repeats was apparently interrupted at the 3' junction. Several points about this series of constructs are worth noting. First, removal of the Clal restriction fragment breaking in EGF repeats 9 and 17 (construct #8 ΔEGF9-17) abolished aggregation with Delta, while reinsertion of this piece into construct #3 ΔCla, which lacks EGF repeats 7-30, restored aggregation to roughly wild type levels (construct #13 ΔCla+EGF9-17), suggesting that EGF repeats 9 through 17 contain sequences important for binding Delta. Second, all constructs in this series (#4-14) were consistent with the ding site mapping to EGF repeats 9 through 17. Expression constructs containing diese repeats (#6. 7, 9, 10. 13) promoted Notch-Delta interactions while constructs lacking these repeats (#4, 5, 8, 1 1. 12, 14) did not. To confirm at inability to aggregate with Delta cells was not simply due to failure of die mutagenized Notch protein to reach the cell surface, but actually reflected the deletion of die necessary binding site, cell surface expression of all constructs was tested by immunofluorescently staining live transfected cells with antibodies specific to die extracellular domain of Notch. All constructs failing to mediate Notch-Delta interactions produced a protein diat appeared to be expressed normally at the cell surface. Third, although the aggregation assay is not quantitative, two constructs which contained EGF repeats 9-17, #9 ΔEGF17-26 or most noticeably #10 ΔEGF26-30, aggregated at a seemingly lower level. Cells transfected with constructs #9 ΔEGF 17-26 and 10 ΔEGF26-30 showed considerably less surface staining dian normal, although fixed and permeabilized cells reacted widi die same antibody stained normally, indicating die epitopes recognized by the antisera had not been simply deleted. By comparing the percentage of transfected cells in eidier permeabilized or live cell populations, it was found that roughly 50% of transfected cells for construct #9 ΔEGF17-26 and 10% for construct #10 ΔEGF26-30 produced detectable protein at the cell surface. Thus diese two constructs produced proteins which often failed to reach the cell surface, perhaps because of misfolding, thereby reducing, but not abolishing, the ability of transfected cells to aggregate with Delta-expressing cells. Having mapped the binding site to EGF repeats 9 through 17, further experiments (Rebay et al. , 1991, Cell 67:687-699) revealed that EGF repeat 14 of Notch was not involved in the interactions with Delta modelled by die tissue culture assay. To further map the Delta binding domain within EGF repeats 9- 17, specific oligonucleotide primers and the PCR technique were used to generate several subfragments of this region. Three overlapping constructs, #16, 17 and 18 were produced, only one of which, #16 ΔCla+EGF9-13, when transfected into S2 cells, allowed aggregation with Delta cells. Construct #19 ΔCla+EGF(10-13), which lacks EGF repeat 9, further defined EGF repeats 10- 13 as die region necessary for Notch-Delta interactions. Constructs #20-24 represented attempts to break this domain down even further using the same PCR strategy (see Figure 3). Constructs #20 ΔCla+EGF(ll-13), in which EGF repeat 12 is the only entire repeat added, and #21 ΔCla+EGF(10-12), in which EGF repeat 11 is the only entire repeat added, failed to mediate aggregation, suggesting that the presence of either EGF repeat 11 or 12 alone was not sufficient for Notch-Delta interactions. However, since the 3' ligation juncture of these constructs interrupted the overall structure of the EGF repeats, it was possible that a short "buffer" zone was needed to allow die crucial repeat to function normally. Thus for example in construct #19 ΔCla+EGF(10-13), EGF repeat 12 might not be directly involved in binding to Delta but instead might contribute the minimum amount of buffer sequence needed to protect die structure of EGF repeat 11 , thereby allowing interactions widi Delta. Constructs #22-24 addressed this issue. Constructs #22 ΔCla+EGF(10-ll), which did not mediate aggregation, and #23 ΔCla+EGF(10- 12), which did, again suggested that both repeats 11 and 12 are required while the flanking sequence from repeat 13 clearly is not. Finally, construct #24 ΔCla+EGF(ll-12), although now potentially structurally disrupted at the 5' junction, convincingly demonstrated that the sequences from EGF repeat 10 are not crucial. Thus based on entirely consistent data from 24 constructs, EGF repeats 11 and 12 of Notch together define the smallest functional unit obtainable from mis analysis that contains the necessary sites for binding to Delta in transfected S2 cells. 7.2.2. EGF REPEATS 11 AND 12 OF NOTCH ARE SUFFICIENT FOR NOTCH-DELTA MEDIATED AGGREGATION The large Clal deletion into which PCR fragments were inserted (#3 ΔCla) retains roughly 1/3 of me original 36 EGF repeats as well as the three Notch/lin-12 repeats. While these are clearly not sufficient to promote aggregation, it is possible that they form a necessary framework widiin which specific EGF repeats can interact with Delta. To test whether only a few EGF repeats were in fact sufficient to promote aggregation, two constructs were designed, #25 ΔEGF which deleted all 36 EGF repeats except for the first two- thirds of repeat 1 , and #30 ΔECN which deleted the entire extracellular portion of Notch except for ie first third of EGF repeat 1 and —35 amino acids just before the transmembrane domain. Fragments which had mediated Notch-Delta aggregation in the background of construct #3 ΔCla, when inserted into construct #25 ΔEGF, were again able to promote interactions with Delta (constructs #26- 30). Analogous constructs (#31 ,32) in which the Notch/lin-12 repeats were also absent, again successfully mediated Notch-Delta aggregation. Thus EGF repeats 11 and 12 appear to function as independent modular units which are sufficient to mediate Notch-Delta interactions in S2 cells, even in the absence of most of the extracellular domain of Notch. 7.2.3. EGF REPEATS 11 AND 12 OF NOTCH 5 MAINTAIN THE CALCIUM DEPENDENCE OF NOTCH-DELTA MEDIATED AGGREGATION The ability of cells expressing certain deletion constructs to aggregate widi Delta expressing cells was examined in the presence or absence of Ca ++ ions. The calcium dependence of the interaction was preserved in even die O smallest construct, consistent with the notion that the minimal constructs containing EGF repeats 11 and 12 bind to Delta in a manner similar to that of full length Notch. 7.2.4. THE DELTA BINDING FUNCTION OF EGF REPEATS 11 AND 12 OF NOTCH IS 5 CONSERVED IN THE XENOPUS HOMOLOG OF NOTCH PCR primers based on the Xenopus Notch sequence (Coffman et al., 1990, Science 249, 1438-1441) were used to obtain an — 350 bp fragment from a Xenopus Stage 17 cDNA library that includes EGF repeats 11 and 12 o flanked by half of repeats 10 and 13 on either side. This fragment was cloned into construct #3 ΔCla, and three independent clones were tested for ability to interact with Delta in die cell culture aggregation assay. Two of the clones, #33a&bΔCla-l-XEGF(10-13). when transfected into S2 cells were able to mediate Notch-Delta interactions at a level roughly equivalent to the analogous Drosophila 5 Notch construct #19ΔCla+EGF(10-13), and again in a calcium dependent manner (Table III). However, the third clone #33cΔCla+XEGF(10-13) failed to mediate Notch-Delta interactions although the protein was expressed normally at the cell surface as judged by staining live unpermeabilized cells. Sequence comparison of die Xenopus PCR product in constructs #33a and 33c revealed a missense 0 mutation resulting in a leucine to proline change (amino acid #453, Coffman, et al., 1990, Science 249, 1438-1441) in EGF repeat 11 of construct #33c. Although this residue is not conserved between Drosophila and Xenopus Notch 5 (Figure 8), the introduction of a proline residue might easily disrupt the structure of the EGF repeat, and thus prevent it from interacting properly with Delta. Comparison of the amino acid sequence of EGF repeats 11 and 12 of Drosophila and Xenopus Notch reveals a high degree of amino acid identity, including d e calcium binding consensus sequence (Figure 4, SEQ ID NO: 1 and NO:2). However the level of homology is not strikingly different from that shared between most of the other EGF repeats, which overall exhibit about 50% identity at die amino acid level. This one to one correspondence between the individual EGF repeats of Drosophila and Xenopus Notch, together with the functional conservation of ELR 11 and 12, suggests that the 36 EGF repeats of Notch comprise a tandem area of conserved functional units. 7.3. DISCUSSION An extensive deletion analysis of die extracellular domain of Notch was used to show that the regions <- Notch containing EGF-homologous repeats 11 and 12 are both necessary and sufficient for Notch-Delta-mediated aggregation, and that this Delta binding capability has been conserved in the same two EGF repeats of Xenopus Notch. The finding that the aggregation mapped to EGF repeats 11 and 12 of Notch demonstrates that the EGF repeats of Notch also function as specific protein binding domains. EGF repeats 11 and 12 alone (#32ΔECN+EGF(11-12)) were sufficient to maintain the Ca ++ dependence of Notch-Delta interactions. The various deletion constructs suggest diat ELR 11 and ELR 12 function as a modular unit, independent of the immediate context into which diey are placed. Thus, neither the remaining 34 EGF repeats nor the three Notch/lin- 12 repeats appear necessary to establish a structural framework required for EGF repeats 11 and 12 to function. Interestingly, almost the opposite effect was observed: aldiough the aggregation assay does not measure the strength of e interaction, as the binding site was narrowed down to smaller and smaller fragments, an increase was observed in die ability of ie transfected cells to aggregate with Delta expressing cells, suggesting that the normal flanking EGF sequences actually impede association between the proteins. The remaining 34 EGF repeats may also form modular binding domains for other proteins interacting with Notch at various times during development. The finding diat EGF repeats 11 and 12 of Notch form a discrete Delta binding unit represents die first concrete evidence supporting die idea diat each EGF repeat or small subset of repeats may play a unique role during development, possibly dirough direct interactions widi odier proteins. The homologies seen between the adhesive domain of Delta and Serrate (Figure 5) suggest that the homologous portion of Serrate is "adhesive" in that it mediates binding to other toporythmic proteins (see Section 8, infra). In addition, the gene scabrous, which encodes a secreted protein with similarity to fibrinogen, may interact with Notch. In addition to the EGF repeat, multiple copies of other structural motifs commonly occur in a variety of proteins. One relevant example is the cdclO/ankyrin motif, six copies of which are found in the intracellular domain of Notch. Ankyrin contains 22 of these repeats. Perhaps repeated arrays of structural motifs may in general represent a linear assembly of a series of modular protein binding units. Given these results together with die known structural, genetic and developmental complexity of Notch. Notch may interact with a number of different ligands in a precisely regulated temporal and spacial pattern d roughout development. Such context specific interactions widi extracellular proteins could be mediated by the EGF and Notch/lin-12 repeats, while interactions with cytoskeletal and cytoplasmic proteins could be mediated by die intracellular cdclO/ankyrin motifs. 8. SEQUENCES WHICH MEDIATE NOTCH-SERRATE INTERACTIONS As described herein, the two EGF repeats of Notch which mediate interactions with Delta, namely EGF repeats 11 and 12, also constitute a Serrate binding domain (see Rebay et al. , 1991 , Cell 67:687-699). To test whether Notch and Serrate directly interact, S2 cells were transfected with a Serrate expression construct and mixed widi Notch expressing cells in an aggregation assay. For the Serrate expression construct, a synthetic primer containing an artificial BamHI site immediately 5' to the initiator AUG at position 442 (all sequence numbers are according to Fleming et al., 1990, Genes & Dev. 4:2188-2201) and homologous through position 464, was used in conjunction with a second primer from position 681-698 to generate a DNA fragment of —260 base pairs. This fragment was cut with BamHI and Kpnl (position 571) and ligated into Bluescript KS-h (Stratagene). This construct, BTSer5'PCR. was checked by sequencing, dien cut with Kpnl. The Serrate Kpnl fragment (571 - 2981) was inserted and the proper orientation selected, to generate BTSer5'PCR-Kpn. The 5' SacII fragment of BTSer5'PCR-Kpn (SacII sites in Bluescript polylinker and in Serrate (1199)) was isolated and used to replace the 5' SacII fragment of cDNA Cl (Fleming et al. , 1990, Genes & Dev. 4:2188-2201), thus regenerating the full length Serrate cDNA minus the 5' untranslated regions. This insert was isolated by a Sail and partial BamHI digestion and shuttled into the BamHI and Sail sites of pRmHa-3 to generate the final expression construct, Ser-mtn. Serrate expressing cells adhered to Notch expressing cells in a calcium dependent manner (Figure 2 and Rebay et al., 1991, supra). However, unlike Delta, under the experimental conditions tested. Serrate did not appear to interact homotypically. In addition, no interactions were detected between Serrate and Delta. A subset of Notch deletion constructs were tested, and showed diat EGF repeats 11 and 12, in addition to binding to Delta, also mediate interactions with Serrate (Figure 2; Constructs #1, 7-10, 13, 16, 17, 19, 28, and 32). In addition, the Serrate-binding function of these repeats also appears to have been conserved in the corresponding two EGF repeats of Xenopus Notch (#33ΔCla+XEGF(10-13)). These results unambiguously show that Notch interacts with bodi Delta and Serrate, and that die same two EGF repeats of Notch mediate both interactions. The Serrate region which is essential for die Notch/Serrate aggregation was also defined. Deleting nucleotides 676-1287 (i.e. amino acids 79-282) (See Figure 5; SEQ ID NO:3 and NO:4) eliminates the ability of the Serrate protein to aggregate widi Notch. Notch and Serrate appear to aggregate less efficiently dian Notch and Delta, perhaps because the Notch-Serrate interaction is weaker. One trivial explanation for this reduced amount of aggregation could be diat the Serrate construct simply did not express as much protein at the cell surface as the Delta construct, thereby diminishing the strength of the interaction. Alternatively, the difference in strength of interaction may indicate a fundamental functional difference between Notch-Delta and Notch-Serrate interactions that may be significant in vivo. 9. THE CLONING, SEQUENCING, AND EXPRESSION OF HUMAN NOTCH 9.1. ISOLATION AND SEQUENCING OF HUMAN NOTCH Clones for the human Notch sequence were originally obtained using the polymerase chain reaction (PCR) to amplify DNA from a 17-18 week human fetal brain cDNA library in the Lambda Zap II vector (Stratagene). The 400 v bp fragment obtained in this manner was then used as a probe widi which to screen the same library for human Notch clones. The original screen yielded three unique clones, hN3k, hN2K, and hN5k, all of which were shown by subsequent sequence analysis to fall in the 3' end of human Notch (Figure 6). A second screen using the 5' end of hN3k as probe was undertaken to search for clones encompassing the 5' end of human Notch. One unique clone, hN4k, was obtained from this screen, and preliminary sequencing data indicate diat it contains most of the 5' end of the gene (Figure 6). Together, clones hN4k, hN3k and hN5k encompass about 10 kb of the human Notch homolog(s), beginning early in the EGF-repeats and extending into the 3' untranslated region of the gene. All three clones are cDNA inserts in the EcoRI site of pBluescript SK ~ (Stratagene). The host E. coU strain is XLl-Blue (see Maniatis, T., 1990, Molecular Cloning, A Laboratory Manual, 2d ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, New York. p. A12). An alignment of the human Notch sequences with Drosophila Notch is shown in Figure 7. The sequence of various portions of Notch contained in the cDNA clones was determined (by use of Sequenase ® , U.S. Biochemical Coφ.) and is shown for hN2k and hN4k in Figures 8 (SEQ ID NO:5-7) and 9 (SEQ ID NO: 8, 9), respectively. Further sequence analysis of hN2k revealed that it encodes a human Notch sequence overlapping diat contained in hN5k. The complete nucleotide sequences of the human Notch cDNA contained in hN3k and hN5k was determined by the dideoxy chain termination method using the Sequenase ® kit (U.S. Biochemical Coφ.). Those nucleotide sequences encoding human Notch, in the appropriate reading frame, were readily identified since diere are no introns and translation in only one out of die diree possible reading frames yields a sequence which, upon comparison with the published Drosophila Notch deduced amino acid sequence, yields a sequence widi a substantial degree of homology to die Drosophila Notch sequence. The DNA and deduced protein sequences of die human Notch cDNA in hN3k and hN5k are presented in Figures 10 (SEQ ID NO: 10. 11) and 11 (SEQ ID NO: 12, 13), respectively. Clone hN3k encodes a portion of a Notch polypeptide starting at approximately the third Notch/lin-12 repeat to several amino acids short of the carboxy-terminal amino acid. Clone hN5k encodes a portion of a Notch polypeptide starting approximately before the cdclO region through the end of the polypeptide, and also contains a 3' untranslated region. Comparing the DNA and protein sequences presented in Figure 10 (SEQ ID NO: 10, 11) with those in Figure 11 (SEQ ID NO: 12, 13) reveals significant differences between the sequences, suggesting that hN3k and hN5k represent part of two distinct Notch-homologous genes. The data thus suggest diat the human genome harbors more than one Notch-homologous gene. This is unlike Drosophila, where Notch appears to be a single-copy gene. Comparison of the DNA and amino acid sequences of the human Notch homologs contained in hN3k and hN5k with the corresponding Drosophila Notch sequences (as published in Wharton et al. , 1985, Cell 43:567-581) and with die corresponding Xenopus Notch sequences (as published in Coffman et al. , 1990, Science 249: 1438-1441 or available from Genbank ® (accession number M33874)) also revealed differences. The amino acid sequence shown in Figure 10 (hN3k) was compared widi the predicted sequence of die TAN-1 polypeptide shown in Figure 2 of Ellisen et al., August 1991, Cell 66:649-661. Some differences were found between the deduced amino acid sequences; however, overall die hN3k Notch polypeptide sequence is 99% identical to die corresponding TAN-1 region (TAN- 1 amino acids 1455 to 2506). Four differences were noted: in the region between the third Notch /l in- 12 repeat and the first cdclO motif, there is an arginine (hN3k) instead of an X (TAN-1 amino acid 1763); (2) there is a proline (hN3k) instead of an X (TAN-1 , amino acid 1787); (3) there is a conservative change of an aspartic acid residue (hN3k) instead of a glutamic acid residue (TAN-1, amino acid 2495); and (4) the carboxyl-terminal region differs substantially between TAN-1 amino acids 2507 and 2535. The amino acid sequence shown in Figure 11 (hN5k) was compared widi die predicted sequence of the TAN-1 polypeptide shown in Figure 2 of Ellisen et al., August 1991 , Cell 66:649-661. Differences were found between the deduced amino acid sequences. The deduced Notch polypeptide of hN5k is 79% identical to the TAN-1 polypeptide (64% identical to Drosophila Notch) in the cdclO region that encompasses both the cclO motif (TAN-1 amino acids 1860 to 2217) and the well-conserved flanking regions (Fig. 12). The cdclO region covers amino acids 1860 through 2217 of the TAN-1 sequence. In addition, the hN5k encoded polypeptide is 65% identical to the TAN-1 polypeptide (44% identical to Drosophila Notch) at the carboxy-terminal end of die molecule containing a PEST (proline, glutamic acid, serine, threonine)-rich region (TAN-1 amino acids 2482 to 2551) (Fig. 12B). The stretch of 215 amino acids lying between the aforementioned regions is not well conserved among any of the Notch-homologous clones represented by hN3k, hN5k, and TAN-1. Neidier die hN5k polypeptide nor Drosophila Notch shows significant levels of amino acid identity to the other proteins in this region (e.g. , hN5k/TAN-l = 24% identity; hN5k/ Drosophila Notch = 11 % identity; TAN-1 /Drosophila Notch = 17% identity). In contrast, Xenopus Notch (Xotch) (SEQ ID NO: 16), rat Notch (SEQ ID NO: 17), and TAN-1 (SEQ ID NO: 18) continue to share significant levels of sequence identity with one anod er (e.g. , TAN- 1 /rat Notch = 75% identity, TAN-l /Xenopus Notch = 45% identity, rat Notch/Xenopus Notch = 50% identity). Examination of the sequence of the intracellular domains of the vertebrate Notch homologs shown in Figure 12B revealed an unexpected finding: all of these proteins, including hN5k, contain a putative CcN motif, associated widi nuclear targeting function, in the conserved region following the last of the six cdclO repeats (Fig. 12B). Although Drosophila Notch lacks such a defined motif, closer inspection of its sequence revealed the presence of a possible bipartite nuclear localization sequence (Robbins et al. , 1991 , Cell 64:615-623), as well as of possible CK II and cdc2 phosphorylation sites, all in relative proximity to one another, thus possibly defining an alternative type of CcN motif (Fig. 12B). To isolate clones covering the 5' end of hN (the human Notch homolog contained in part in hN5k), clone hN2k was used as a probe to screen 260,000 plaques of human fetal brain phage library, commercially available from Stratagene, for crosshybridizing clones. Four clones were identified and isolated using standard procedures (Maniatis et al. , 1982, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York). Four clones were also isolated by hybridization to die Notch-homologous sequence of Adams et al., 1992, Nature 355:632-655, which was obtained from the ATCC. To isolate clones covering the 5' end of TAN- 1. the human fetal brain library that is commercially available from Stratagene was screened for clones which would extend the sequence to the 5' end. 880.000 plaques were screened and four clones were identified which crosshybridized with die hN3k sequences. Sequencing confirmed die relative position of diese sequences wi in the Notch protein encoded by TAN-1. The 5' sequence of our isolated TAN-1 homolog has been determined dirough nucleotide number 972 (nucleotide number 1 being the A in the ATG initiation codon), and compared to the sequence as published by Ellisen et- al (1991 , Cell 66:649-661). At nucleotide 559, our TAN-1 homolog has a G, whereas Ellisen et al. disclose an A, which change results in a different encoded amino acid. Thus, within the first 324 amino acids, our TAN-1-encoded protein differs from that taught by Ellisen et al., since our protein has a Gly at position 187, whereas Ellisen et al. disclose an Arg at diat position (as presented in Figure 13.) The full-length amino acid sequences of both the hN (SEQ ID NO: 19) and TAN-1-encoded (SEQ ID NO:20) proteins, as well as Xenopus and Drosophila Notch proteins, are shown in Figure 13. The full-length DNA coding sequence (except for that encoding the initiator Met) (contained in SEQ ID NO:21) and encoded amino acid sequence (except that the initiator Met is not shown) (contained in SEQ ID NO: 19) of hN are shown in Figure 17. 9.2. EXPRESSION OF HUMAN NOTCH Expression constructs were made using die human Notch cDNA clones discussed in Section 9.1 above. In the cases of hN3k and hN2k, the entire clone was excised from its vector as an EcoRI restriction fragment and subcloned into e EcoRI restriction site of each of the three pGEX vectors (Glutathione S- Transferase expression vectors; Smith and Johnson, 1988, Gene 7, 31-40). This allows for the expression of the Notch protein product from die subclone in the correct reading frame. In the case of hN5k, the clone contains two internal EcoRI restriction sites, producing 2.6, 1.5 and 0.6 kb fragments. Both the 2.6 and the 1.5 kb fragments have also been subcloned into each of the pGEX vectors. The pGEX vector system was used to obtain expression of human Notch fusion (chimeric) proteins from the constructs described below. The cloned Notch DNA in each case was inserted, in phase, into die appropriate pGEX vector. Each construct was then electroporated into bacteria (E. coli). and was expressed as a fusion protein containing die Notch protein sequences fused to the carboxyl terminus of glutathione S-transferase protein. Expression of the fusion proteins was confirmed by analysis of bacterial protein extracts by polyacrylamide gel electrophoresis, comparing protein extracts obtained from bacteria containing the pGEX plasmids with and without the inserted Notch DNA. The fusion proteins were soluble in aqueous solution, and were purified from bacterial lysates by affinity chromatography using glutathione-coated agarose (since the carboxyl terminus of glutathione S-transferase binds to glutadiionine). The expressed fusion proteins were bound by an antibody to Drosophila Notch, as assayed by Western blotting. The constructs used to make human Notch-glutathione S- transferase fusion proteins were as follows: hNFP#2 - PCR was used to obtain a fragment starting just before the cdclO repeats at nucleotide 192 of die hN5k insert to just before the PEST-rich region at nucleotide 1694. The DNA was then digested with BamHI and Smal and the resulting fragment was ligated into pGEX-3. After expression, die fusion protein was purified by binding to glutadiione agarose. The purified polypeptide was quantitated on a 4-15% gradient polyacrylamide gel. The resulting fusion protein had an approximate molecular weight of 83 kD. hN3FP#l - The entire hN3k DNA insert (nucleotide 1 to 3235) was excised from the Bluescript (SK) vector by digesting with EcoRI. The DNA was ligated into pGEX-3. hN3FP#2 - A 3' segment of hN3k DNA (nucleotide 1847 to 3235) plus some of the polylinker was cut out of the Bluescript (SK) vector by digesting with Xmal. The fragment was ligated into pGEX-1. Following purification, these fusion proteins are used to make either polyclonal and/or monoclonal antibodies to human Notch. 10. NOTCH EXPRESSION IN NORMAL AND MALIGNANT CELLS Various human patient tissue samples and cell lines, representing both normal and a wide variety of malignant cells are assayed to detect and/or 5 quantitate expression of Notch. Patient tissue samples are obtained from the pathology department at die Yale University School of Medicine. The following assays are used to measure Notch expression in patient tissue samples: (a) Northern hybridization; (b) Western blots; (c) in situ hybridization; and (d) immunocytochemistry. Assays are carried out using O standard techniques. Northern hybridization and in situ hybridization are carried out (i) using a DNA probe specific to die Notch sequence of clone hN3k; and (ii) using a DNA probe specific to the Notch sequence of clone hN5k. Western blots and immunocytochemistry are carried out using an antibody to Drosophila Notch protein (which also recognizes human Notch proteins). J Northern hybridization and Western blots, as described above, are also used to analyze numerous human cell lines, representing various normal or cancerous tissues. The cell lines tested are listed in Table 2. 20 Table 2 HUMAN CELL LINES 5 30 35 Colon Caco-2 SW 48 T84 WiDr Embryo FHs 173 We Kidney A-498 A-704 Caki-2 Leukemia ARH-77 KG-1 Liver Hep G2 WRL 68 Lung Calu-1 HLF-a SK-Lu-1 Lymphoblasts CCRF-CEM HuT 78 Lymphoma Hs 445 MS116 U-937 Melanoma A-375 G-361 Hs 294T SK-MEL-1 Myeloma IM-9 RPMI 8226 Neuroblastoma 1MR-32 SK-N-SH SK-N-MC Ovary Caov-3 Caov-4 PA-1 Plasma Cells ARH-77 Sarcoma A-204 A673 HOS Skin Amdur II BUD-8 Testis Tera-1 Tera-2 Thymus Hs67 Uterus AN3 Ca HEC-l-A Malignancies of malignant cell tissue types which are thus shown to specifically express Notch can be treated as described in Section 5.1 et seq. 10.1. EXPRESSION OF HUMAN NOTCH PROTEIN IS INCREASED IN VARIOUS MALIGNANCIES As described below, we have found that human Notch protein expression is increased in at least three human cancers, namely cervical, breast, and colon cancer. Immunocytochemical staining of tissue samples from cervical, breast, and colon cancers of human patients showed clearly that the malignant tissue expresses high levels of Notch, at increased levels relative to non-malignant tissue sections. This broad spectrum of different neoplasias in which diere is elevated Notch expression suggests diat many more cancerous conditions will be seen to upregulate Notch. Slides of human tumor samples (for breast, colon, and cervical tumors) were obtained from the tissue bank of the Pathology Department, Yale Medical School. The stainings were done using monoclonal antibodies raised against the PI and P4 fusion proteins which were generated from sequences of hN and TAN-1, respectively. The PI and P4 fusion proteins were obtained by insertion of the desired human Notch sequence into the appropriate pGEX expression vector (Smith and Johnson, 1988, Gene 7:31-40; AMRAD Coφ. , Melbourne, Australia) and were affinity-purified according to the instructions of the manufacturer (AMRAD Coφ.). For production of the PI fusion protein, pGEX-2 was cut with BamHI and ligated to a concatamer which consists of three copies of a 518 bp BamHI-Bglll fragment of hN. Rats were immunized with die expressed protein and monoclonal antibodies were produced by standard procedures. For production of die P4 fusion protein, pGEX-2 was cut widi BamHI and ligated to a concatamer which consists of three copies of a 473 bp BamHI-Bglll fragment of TAN-1. Rats were immunized widi the expressed protein, and monoclonal antibodies were produced by standard procedures. In all tumors examined, the Notch proteins encoded by both human Notch homologs TAN-1 and hN were present at increased levels in die malignant part of the tissue compared to the normal part. Representative stainings are shown in the pictures provided (Figs. 14-16). The staining procedure was as follows: The tissues were fixed in paraformaldehyde, embedded in paraffin, cut in 5 micrometer thick sections and placed on glass slides. Then the following steps were carried out: 1. Deparafinization dirough 4 changes of xylene, 4 minutes each. 2. Removal of xylene through 3 changes in absolute ethanol, 4 minutes each. 3. Gradual rehydration of die tissues by immersing d e slides into 95%, 90%, 80%, 60% and 30% ethanol, 4 minutes each. At the end die slides were rinsed in distilled water for 5 minutes. 4. Quenching of endogenous, peroxidase by incubating for 30 minutes in 0.3% hydrogen peroxide in methanol. 5. Washing in PBS (10 mM sodium phosphate pH 7.5, 0.9% NaCl) for 20 minutes. 6. Incubation for 1 hour in blocking solution. (Blocking solution: PBS containing 4% normal rabbit serum and 0.1 Triton X-100.) 88 - 11. DEPOSIT OF MICROORGANISMS The following recombinant bacteria, each carrying a plasmid encoding a portion of human Notch, were deposited on May 2, 1991 widi die American Type Culture Collection, 1201 Parklawn Drive, Rockville, Maryland 20852, under die provisions of the Budapest Treaty on die International Recognition of the Deposit of Microorganisms for the Puφoses of Patent Procedures. Bacteria carrying Plasmid ATCC Accession No. E. coli XLl-Blue hN4k 68610 E. coH XLl-Blue hN3k 68609 E. coli XLl-Blue hN5k 68611 The present invention is not to be limited in scope by the microorganisms deposited or the specific embodiments described herein. Indeed, various modifications of the invention in addition to diose described herein will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall widiin die scope of the appended claims. Various publications are cited herein, the disclosures of which are incoφorated by reference in their entireties. - 87 - 7. Incubation overnight at 4°C with primary antibody diluted in blocking solution. Final concentration of primary antibody 20-50 μg/ml. 8. Washing for 20 minutes with PBS+0.1 % Triton X-100 (3 changes). 9. Incubation for 30 minutes with biotinylated rabbit anti-rat antibody: 50 μl of biotinylated antibody (VECTOR) in 10 ml of blocking solution. 10. Washing for 20 minutes with PBS +0.1 % Triton X-100 (3 changes). 11. Incubation with ABC reagent (VECTOR) for 30 minutes (the reagent is made in PBS+0.1 % Triton X-100). 12. Washing for 20 minutes in PBS+0.1 % Triton X-100. Followed by incubation for 2 minutes in PBS +0.5% Triton X-100. 13. Incubation for 2-5 minutes in peroxidase substrate solution. Peroxidase substrate solution: Equal volumes of 0.02% hydrogen peroxide in distilled water and 0.1 % diaminobenzidine tetrahydrochloride (DAB) in 0.1 M Tris buffer pH 7.5 are mixed just before the incubation with the tissues. Triton X-100 is added to the final solution at a concentration of 0.5%. 14. Washing for 15 minutes in tap water. 15. Counterstaining for 10 minutes with Mayer's hematoxylin. 16. Washing for 15 minutes in tap water. 17. Dehydration through changes in 30%, 60%, 80%, 90%, 95% and absolute edianol (4 minutes each). 18. Immersion into xylene (2 changes, 4 minutes each). 19. Mounting, light microscopy. 88.1 - International Application No: PCT/ / MICROORGANISMS Optional Sheet in connection with the microorganism referred to on page 88, lines 1-12 of the description ' A. IDENTIFICATION Of DEPOSIT ' Further deposits are identified on an additional sheet ' Name of depositary institution ' American Type Culture Collection Address of depositary institution (including postal code and country) ' 12301 Parklawn Drive Rockville. MD 20852 US Date of deposit ' May 2. 1991 Accession Number ' 68610 B. ADDITIONAL INDICATIONS ' (love blank if not applicable). This information u continued on a separate attached sheet C. DESIGNATED STATES FOR WHICH INDICATIONS ARE MADE ' «.__.__ D. SEPARATE FURNISHING OF INDICATIONS * flew, blank if no. ap licable) The indications listed below wilt be submitted to the International Bureeu later ' (Specify the general nature of the indications e.g., "Accession Number of Deposit*) E. B"This sheet was received with the International application when filed (to be checked by the receiving Office) (Authorized Officer) D The date of receipt (from the applicant) by the International Bureau " (Authorized Officer) Form PCT/RO/134 (January 1981 ) 88.2 International Application No: PCT/ Form PCT/RO/134 (cont.) American Type Culture Collection 12301 Parklawn Drive Rockville, MD 20852 US Accession No. Date of Deposit 68609 May 2, 1991 6861 1 May 2, 1991 -89- SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Artavanis-Tsakonas, S. et al. (ii) TITLE OF INVENTION: Therapeutic And Diagnostic Methods And Compositions Based On Notch Proteins And Nucleic Acids (iii) NUMBER OF SEQUENCES: 21 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: Pennie ~ Edmonds (B) STREET: 1155 Avenue of the Americas (C) CITY: New York (D) STATE: New York (E) COUNTRY: U.S.A. (F) ZIP: 10036 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: PatentIn Release #1.0, Version #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: To be assigned (B) FILING DATE: On even date (C) CLASSIFICATION: (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Misrock, S. Leslie (B) REGISTRATION NUMBER: 18,872 (C) REFERENCE/DOCKET NUMBER: 7326-018 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: 212 790-9090 (B) TELEFAX: 212 8698864/9741 (C) TELEX: 66141 PENNIE (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2892 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: cDNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 142..2640 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: GAATTCGGAG GAATTATTCA AAACATAAAC ACAATAAACA ATTTGAGTAG TTGCCGCACA 60 CACACACACA CACAGCCCGT GGATTATTAC ACTAAAAGCG ACACTCAATC CAAAAAATCA 120 GCAACAAAAA CATCAATAAA C ATG CAT TGG ATT AAA TGT TTA TTA ACA GCA 171 -90- Met His Trp lie Lys Cys Leu Leu Thr Ala 1 5 10 TTC ATT TGC TTC ACA GTC ATC GTG CAG GTT CAC AGT TCC GGC AGC TTT 219 Phe lie Cys Phe Thr Val lie Val Gin Val His Ser Ser Gly Ser Phe 15 20 25 GAG TTG CGC CTG AAG TAC TTC AGC AAC GAT CAC GGG CGG GAC AAC GAG 267 Glu Leu Arg Leu Lys Tyr Phe Ser Asn Asp His Gly Arg Asp Asn Glu 30 35 40 GGT CGC TGC TGC AGC GGG GAG TCG GAC GGA GCG ACG GGC AAG TGC CTG 315 Gly Arg Cys Cys Ser Gly Glu Ser Asp Gly Ala Thr Gly Lys Cys Leu 45 50 55 GGC AGC TGC AAG ACG CGG TTT CGC GTC TGC CTA AAG CAC TAC CAG GCC 363 Gly Ser Cys Lys Thr Arg Phe Arg Val Cys Leu Lys His Tyr Gin Ala 60 65 70 ACC ATC GAC ACC ACC TCC CAG TGC ACC TAC GGG GAC GTG ATC ACG CCC 411 Thr lie Asp Thr Thr Ser Gin Cys Thr Tyr Gly Asp Val lie Thr Pro 75 80 85 90 ATT CTC GGC GAG AAC TCG GTC AAT CTG ACC GAC GCC CAG CGC TTC CAG 459 lie Leu Gly Glu Asn Ser Val Asn Leu Thr Asp Ala Gin Arg Phe Gin 95 100 105 AAC AAG GGC TTC ACG AAT CCC ATC CAG TTC CCC TTC TCG TTC TCA TGG 507 Asn Lys Gly Phe Thr Asn Pro lie Gin Phe Pro Phe Ser Phe Ser Trp 110 115 120 CCG GGT ACC TTC TCG CTG ATC GTC GAG GCC TGG CAT GAT ACG AAC AAT 555 Pro Gly Thr Phe Ser Leu lie Val Glu Ala Trp His Asp Thr Asn Asn 125 130 135 AGC GGC AAT GCG CGA ACC AAC AAG CTC CTC ATC CAG CGA CTC TTG GTG 603 Ser Gly Asn Ala Arg Thr Asn Lys Leu Leu lie Gin Arg Leu Leu Val 140 145 150 CAG CAG GTA CTG GAG GTG TCC TCC GAA TGG AAG ACG AAC AAG TCG GAA 651 Gin Gin Val Leu Glu Val Ser Ser Glu Trp Lys Thr Asn Lys Ser Glu 155 160 165 170 TCG CAG TAC ACG TCG CTG GAG TAC GAT TTC CGT GTC ACC TGC GAT CTC 699 Ser Gin Tyr Thr Ser Leu Glu Tyr Asp Phe Arg Val Thr Cys Asp Leu 175 180 185 AAC TAC TAC GGA TCC GGC TGT GCC AAG TTC TGC CGG CCC CGC GAC GAT 747 Asn Tyr Tyr Gly Ser Gly Cys Ala Lys Phe Cys Arg Pro Arg Asp Asp 190 195 200 TCA TTT GGA CAC TCG ACT TGC TCG GAG ACG GGC GAA ATT ATC TGT TTG 795 Ser Phe Gly His Ser Thr Cys Ser Glu Thr Gly Glu lie lie Cys Leu 205 210 215 ACC GGA TGG CAG GGC GAT TAC TGT CAC ATA CCC AAA TGC GCC AAA GGC 843 Thr Gly Trp Gin Gly Asp Tyr Cys His lie Pro Lys Cys Ala Lys Gly 220 225 230 TGT GAA CAT GGA CAT TGC GAC AAA CCC AAT CAA TGC GTT TGC CAA CTG 891 Cys Glu His Gly His Cys Asp Lys Pro Asn Gin Cys Val Cys Gin Leu 235 240 245 250 GGC TGG AAG GGA GCC TTG TGC AAC GAG TGC GTT CTG GAA CCG AAC TGC 939 Gly Trp Lys Gly Ala Leu Cys Asn Glu Cys Val Leu Glu Pro Asn Cys 255 260 265 -91- ATC CAT GGC ACC TGC AAC AAA CCC TGG ACT TGC ATC TGC AAC GAG GGT 987 lie His Gly Thr Cys Asn Lys Pro Trp Thr Cys lie Cys Asn Glu Gly 270 275 280 TGG GGA GGC TTG TAC TGC AAC CAG GAT CTG AAC TAC TGC ACC AAC CAC 1035 Trp Gly Gly Leu Tyr Cys Asn Gin Asp Leu Asn Tyr Cys Thr Asn His 285 290 295 AGA CCC TGC AAG AAT GGC GGA ACC TGC TTC AAC ACC GGC GAG GGA TTG 1083 Arg Pro Cys Lys Asn Gly Gly Thr Cys Phe Asn Thr Gly Glu Gly Leu 300 305 310 TAC ACA TGC AAA TGC GCT CCA GGA TAC AGT GGT GAT GAT TGC GAA AAT 1131 Tyr Thr Cys Lys Cys Ala Pro Gly Tyr Ser Gly Asp Asp Cys Glu Asn 315 320 325 330 GAG ATC TAC TCC TGC GAT GCC GAT GTC AAT CCC TGC CAG AAT GGT GGT 1179 Glu lie Tyr Ser Cys Asp Ala Asp Val Asn Pro Cys Gin Asn Gly Gly 335 340 345 ACC TGC ATC GAT GAG CCG CAC ACA AAA ACC GGC TAC AAG TGT CAT TGC 1227 Thr Cys lie Asp Glu Pro His Thr Lys Thr Gly Tyr Lys Cys His Cys 350 355 360 GCC AAC GGC TGG AGC GGA AAG ATG TGC GAG GAG AAA GTG CTC ACG TGT 1275 Ala Asn Gly Trp Ser Gly Lys Met Cys Glu Glu Lys Val Leu Thr Cys 365 370 375 TCG GAC AAA CCC TGT CAT CAG GGA ATC TGC CGC AAC GTT CGT CCT GGC 1323 Ser Asp Lys Pro Cys His Gin Gly lie Cys Arg Asn Val Arg Pro Gly 380 385 390 TTG GGA AGC AAG GGT CAG GGC TAC CAG TGC GAA TGT CCC ATT GGC TAC 1371 Leu Gly Ser Lys Gly Gin Gly Tyr Gin Cys Glu Cys Pro lie Gly Tyr 395 400 405 410 AGC GGA CCC AAC TGC GAT CTC CAG CTG GAC AAC TGC AGT CCG AAT CCA 1419 Ser Gly Pro Asn Cys Asp Leu Gin Leu Asp Asn Cys Ser Pro Asn Pro 415 420 425 TGC ATA AAC GGT GGA AGC TGT CAG CCG AGC GGA AAG TGT ATT TGC CCA 1467 Cys lie Asn Gly Gly Ser Cys Gin Pro Ser Gly Lys Cys lie Cys Pro 430 435 440 GCG GGA TTT TCG GGA ACG AGA TGC GAG ACC AAC ATT GAC GAT TGT CTT 1515 Ala Gly Phe Ser Gly Thr Arg Cys Glu Thr Asn lie Asp Asp Cys Leu 445 450 455 GGC CAC CAG TGC GAG AAC GGA GGC ACC TGC ATA GAT ATG GTC AAC CAA 1563 Gly His Gin Cys Glu Asn Gly Gly Thr Cys lie Asp Met Val Asn Gin 460 465 470 TAT CGC TGC CAA TGC GTT CCC GGT TTC CAT GGC ACC CAC TGT AGT AGC 1611 Tyr Arg Cys Gin Cys Val Pro Gly Phe His Gly Thr His Cys Ser Ser 475 480 485 490 AAA GTT GAC TTG TGC CTC ATC AGA CCG TGT GCC AAT GGA GGA ACC TGC 1659 Lys Val Asp Leu Cys Leu lie Arg Pro Cys Ala Asn Gly Gly Thr Cys 495 500 505 TTG AAT CTC AAC AAC GAT TAC CAG TGC ACC TGT CGT GCG GGA TTT ACT 1707 Leu Asn Leu Asn Asn Asp Tyr Gin Cys Thr Cys Arg Ala Gly Phe Thr 510 515 520 GGC AAG GAT TGC TCT GTG GAC ATC GAT GAG TGC AGC AGT GGA CCC TGT 1755 Gly Lys Asp Cys Ser Val Asp lie Asp Glu Cys Ser Ser Gly Pro Cys 525 530 535 -92- CAT AAC GGC GGC ACT TGC ATG AAC CGC GTC AAT TCG TTC GAA TGC GTG 1803 His Asn Gly Gly Thr Cys Met Asn Arg Val Asn Ser Phe Glu Cys Val 540 545 550 TGT GCC AAT GGT TTC AGG GGC AAG CAG TGC GAT GAG GAG TCC TAC GAT 1851 Cys Ala Asn Gly Phe Arg Gly Lys Gin Cys Asp Glu Glu Ser Tyr Asp 555 560 565 570 TCG GTG ACC TTC GAT GCC CAC CAA TAT GGA GCG ACC ACA CAA GCG AGA 1899 Ser Val Thr Phe Asp Ala His Gin Tyr Gly Ala Thr Thr Gin Ala Arg 575 580 585 GCC GAT GGT TTG ACC AAT GCC CAG GTA GTC CTA ATT GCT GTT TTC TCC 1947 Ala Asp Gly Leu Thr Asn Ala Gin Val Val Leu lie Ala Val Phe Ser 590 595 600 GTT GCG ATG CCT TTG GTG GCG GTT ATT GCG GCG TGC GTG GTC TTC TGC 1995 Val Ala Met Pro Leu Val Ala Val lie Ala Ala Cys Val Val Phe Cys 605 610 615 ATG AAG CGC AAG CGT AAG CGT GCT CAG GAA AAG GAC GAC GCG GAG GCC 2043 Met Lys Arg Lys Arg Lys Arg Ala Gin Glu Lys Asp Asp Ala Glu Ala 620 625 630 AGG AAG CAG AAC GAA CAG AAT GCG GTG GCC ACA ATG CAT CAC AAT GGC 2091 Arg Lys Gin Asn Glu Gin Asn Ala Val Ala Thr Met His His Asn Gly 635 640 645 650 AGT GGG GTG GGT GTA GCT TTG GCT TCA GCC TCT CTG GGC GGC AAA ACT 2139 Ser Gly Val Gly Val Ala Leu Ala Ser Ala Ser Leu Gly Gly Lys Thr 655 660 665 GGC AGC AAC AGC GGT CTC ACC TTC GAT GGC GGC AAC CCG AAT ATC ATC 2187 Gly Ser Asn Ser Gly Leu Thr Phe Asp Gly Gly Asn Pro Asn lie lie 670 675 680 AAA AAC ACC TGG GAC AAG TCG GTC AAC AAC ATT TGT GCC TCA GCA GCA 2235 Lys Asn Thr Trp Asp Lys \ Ser Val Asn Asn lie Cys Ala Ser Ala Ala 685 690 695 GCA GCG GCG GCG GCG GCA GCA GCG GCG GAC GAG TGT CTC ATG TAC GGC 2283 Ala Ala Ala Ala Ala Ala Ala Ala Ala Asp Glu Cys Leu Met Tyr Gly 700 705 710 GGA TAT GTG GCC TCG GTG GCG GAT AAC AAC AAT GCC AAC TCA GAC TTT 2331 Gly Tyr Val Ala Ser Val Ala Asp Asn Asn Asn Ala Asn Ser Asp Phe 715 720 725 730 TGT GTG GCT CCG CTA CAA AGA GCC AAG TCG CAA AAG CAA CTC AAC ACC 2379 Cys Val Ala Pro Leu Gin Arg Ala Lys Ser Gin Lys Gin Leu Asn Thr 735 740 745 GAT CCC ACG CTC ATG CAC CGC GGT TCG CCG GCA GGC AGC TCA GCC AAG 2427 Asp Pro Thr Leu Met His Arg Gly Ser Pro Ala Gly Ser Ser Ala Lys 750 755 760 GGA GCG TCT GGC GGA GGA CCG GGA GCG GCG GAG GGC AAG AGG ATC TCT 2475 Gly Ala Ser Gly Gly Gly Pro Gly Ala Ala Glu Gly Lys Arg lie Ser 765 770 775 GTT TTA GGC GAG GGT TCC TAC TGT AGC CAG CGT TGG CCC TCG TTG GCG 2523 Val Leu Gly Glu Gly Ser Tyr Cys Ser Gin Arg Trp Pro Ser Leu Ala 780 785 790 GCG GCG GGA GTG GCC GGA GCC TGT TCA TCC CAG CTA ATG GCT GCA GCT 2571 Ala Ala Gly Val Ala Gly Ala Cys Ser Ser Gin Leu Met Ala Ala Ala 795 800 805 810 -93- TCG GCA GCG GGC AGC GGA GCG GGG ACG GCG CAA CAG CAG CGA TCC GTG 2619 Ser Ala Ala Gly Ser Gly Ala Gly Thr Ala Gin Gin Gin Arg Ser Val 815 820 825 GTC TGC GGC ACT CCG CAT ATG TAACTCCAAA AATCCGGAAG GGCTCCTGGT 2670 Val Cys Gly Thr Pro His Met 830 AAATCCGGAG AAATCCGCAT GGAGGAGCTG ACAGCACATA CACAAAGAAA AGACTGGGTT 2730 GGGTTCAAAA TGTGAGAGAG ACGCCAAAAT GTTGTTGTTG ATTGAAGCAG TTTAGTCGTC 2790 ACGAAAAATG AAAAATCTGT AACAGGCATA ACTCGTAAAC TCCCTAAAAA ATTTGTATAG 2850 TAATTAGCAA AGCTGTGACC CAGCCGTTTC GATCCCGAAT TC 2892 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 833 amino acids (B) TYPE: amino acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Met His Trp lie Lys Cys Leu Leu Thr Ala Phe lie Cys Phe Thr Val 1 5 10 15 lie Val Gin Val His Ser Ser Gly Ser Phe Glu Leu Arg Leu Lys Tyr 20 25 30 Phe Ser Asn Asp His Gly Arg Asp Asn Glu Gly Arg Cys Cys Ser Gly 35 40 45 Glu Ser Asp Gly Ala Thr^ Gly Lys Cys Leu Gly Ser Cys Lys Thr Arg 50 55 60 Phe Arg Val Cys Leu Lys His Tyr Gin Ala Thr lie Asp Thr Thr Ser 65 70 75 80 Gin Cys Thr Tyr Gly Asp Val lie Thr Pro lie Leu Gly Glu Asn Ser 85 90 95 Val Asn Leu Thr Asp Ala Gin Arg Phe Gin Asn Lys Gly Phe Thr Asn 100 105 110 Pro lie Gin Phe Pro Phe Ser Phe Ser Trp Pro Gly Thr Phe Ser Leu 115 120 125 lie Val Glu Ala Trp His Asp Thr Asn Asn Ser Gly Asn Ala Arg Thr 130 135 140 Asn Lys Leu Leu lie Gin Arg Leu Leu Val Gin Gin Val Leu Glu Val 145 150 155 160 Ser Ser Glu Trp Lys Thr Asn Lys Ser Glu Ser Gin Tyr Thr Ser Leu 165 170 175 Glu Tyr Asp Phe Arg Val Thr Cys Asp Leu Asn Tyr Tyr Gly Ser Gly 180 185 190 Cys Ala Lys Phe Cys Arg Pro Arg Asp Asp Ser Phe Gly His Ser Thr 195 200 205 -94- Cys Ser Glu Thr Gly Glu lie lie Cys Leu Thr Gly Trp Gin Gly Asp 210 215 220 Tyr Cys His lie Pro Lys Cys Ala Lys Gly Cys Glu His Gly His Cys 225 230 235 240 Asp Lys Pro Asn Gin Cys Val Cys Gin Leu Gly Trp Lys Gly Ala Leu 245 250 255 Cys Asn Glu Cys Val Leu Glu Pro Asn Cys lie His Gly Thr Cys Asn 260 265 270 Lys Pro Trp Thr Cys lie Cys Asn Glu Gly Trp Gly Gly Leu Tyr Cys 275 280 285 Asn Gin Asp Leu Asn Tyr Cys Thr Asn His Arg Pro Cys Lys Asn Gly 290 295 300 Gly Thr Cys Phe Asn Thr Gly Glu Gly Leu Tyr Thr Cys Lys Cys Ala 305 310 315 320 Pro Gly Tyr Ser Gly Asp Asp Cys Glu Asn Glu lie Tyr Ser Cys Asp 325 330 335 Ala Asp Val Asn Pro Cys Gin Asn Gly Gly Thr Cys lie Asp Glu Pro 340 345 350 His Thr Lys Thr Gly Tyr Lys Cys His Cys Ala Asn Gly Trp Ser Gly 355 360 365 Lys Met Cys Glu Glu Lys Val Leu Thr Cys Ser Asp Lys Pro Cys His 370 375 380 Gin Gly lie Cys Arg Asn Val Arg Pro Gly Leu Gly Ser Lys Gly Gin 385 390 395 400 Gly Tyr Gin Cys Glu Cys Pro lie Gly Tyr Ser Gly Pro Asn Cys Asp 405 410 415 Leu Gin Leu Asp Asn Cys Ser Pro Asn Pro Cys lie Asn Gly Gly Ser 420 425 430 Cys Gin Pro Ser Gly Lys Cys lie Cys Pro Ala Gly Phe Ser Gly Thr 435 440 445 Arg Cys Glu Thr Asn lie Asp Asp Cys Leu Gly His Gin Cys Glu Asn 450 455 460 Gly Gly Thr Cys lie Asp Met Val Asn Gin Tyr Arg Cys Gin Cys Val 465 470 475 480 Pro Gly Phe His Gly Thr His Cys Ser Ser Lys Val Asp Leu Cys Leu 485 490 495 lie Arg Pro Cys Ala Asn Gly Gly Thr Cys Leu Asn Leu Asn Asn Asp 500 505 510 Tyr Gin Cys Thr Cys Arg Ala Gly Phe Thr Gly Lys Asp Cys Ser Val 515 520 525 Asp lie Asp Glu Cys Ser Ser Gly Pro Cys His Asn Gly Gly Thr Cys 530 535 540 Met Asn Arg Val Asn Ser Phe Glu Cys Val Cys Ala Asn Gly Phe Arg 545 550 555 560 Gly Lys Gin Cys Asp Glu Glu Ser Tyr Asp Ser Val Thr Phe Asp Ala -95- 565 570 575 His Gin Tyr Gly Ala Thr Thr Gin Ala Arg Ala Asp Gly Leu Thr Asn 580 585 590 Ala Gin Val Val Leu lie Ala Val Phe Ser Val Ala Met Pro Leu Val 595 600 605 Ala Val lie Ala Ala Cys Val Val Phe Cys Met Lys Arg Lys Arg Lys 610 615 620 Arg Ala Gin Glu Lys Asp Asp Ala Glu Ala Arg Lys Gin Asn Glu Gin 625 630 635 640 Asn Ala Val Ala Thr Met His His Asn Gly Ser Gly Val Gly Val Ala 645 650 655 Leu Ala Ser Ala Ser Leu Gly Gly Lys Thr Gly Ser Asn Ser Gly Leu 660 665 670 Thr Phe Asp Gly Gly Asn Pro Asn lie lie Lys Asn Thr Trp Asp Lys 675 680 685 Ser Val Asn Asn lie Cys Ala Ser Ala Ala Ala Ala Ala Ala Ala Ala 690 695 700 Ala Ala Ala Asp Glu Cys Leu Met Tyr Gly Gly Tyr Val Ala Ser Val 705 710 715 720 Ala Asp Asn Asn Asn Ala Asn Ser Asp Phe Cys Val Ala Pro Leu Gin 725 730 735 Arg Ala Lys Ser Gin Lys Gin Leu Asn Thr Asp Pro Thr Leu Met His 740 745 750 Arg Gly Ser Pro Ala Gly Ser Ser Ala Lys Gly Ala Ser Gly Gly Gly 755 760 765 Pro Gly Ala Ala Glu Gly Lys Arg lie Ser Val Leu Gly Glu Gly Ser 770 775 780 Tyr Cys Ser Gin Arg Trp Pro Ser Leu Ala Ala Ala Gly Val Ala Gly 785 790 795 800 Ala Cys Ser Ser Gin Leu Met Ala Ala Ala Ser Ala Ala Gly Ser Gly 805 810 815 Ala Gly Thr Ala Gin Gin Gin Arg Ser Val Val Cys Gly Thr Pro His 820 825 830 Met (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1320 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: cDNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 442..1320 -96- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: CCGAGTCGAG CGCCGTGCTT CGAGCGGTGA TGAGCCCCTT TTCTGTCAAC GCTAAAGATC 60 TACAAAACAT CAGCGCCTAT CAAGTGGAAG TGTCAAGTGT GAACAAAACA AAAACGAGAG 120 AAGCACATAC TAAGGTCCAT ATAAATAATA AATAATAATT GTGTGTGATA ACAACATTAT 180 CCAAACAAAA CCAAACAAAA CGAAGGCAAA GTGGAGAAAA TGATACAGCA TCCAGAGTAC 240 GGCCGTTATT CAGCTATCCA GAGCAAGTGT AGTGTGGCAA AATAGAAACA AACAAAGGCA 300 CCAAAATCTG CATACATGGG CTAATTAAGG CTGCCCAGCG AATTTACATT TGTGTGGTGC 360 CAATCCAGAG TGAATCCGAA ACAAACTCCA TCTAGATCGC CAACCAGCAT CACGCTCGCA 420 AACGCCCCCA GAATGTACAA A ATG TTT AGG AAA CAT TTT CGG CGA AAA CCA 471 Met Phe Arg Lys His Phe Arg Arg Lys Pro 1 5 10 GCT ACG TCG TCG TCG TTG GAG TCA ACA ATA GAA TCA GCA GAC AGC CTG 519 Ala Thr Ser Ser Ser Leu Glu Ser Thr lie Glu Ser Ala Asp Ser Leu 15 20 25 GGA ATG TCC AAG AAG ACG GCG ACA AAA AGG CAG CGT CCG AGG CAT CGG 567 Gly Met Ser Lys Lys Thr Ala Thr Lys Arg Gin Arg Pro Arg His Arg 30 35 40 GTA CCC AAA ATC GCG ACC CTG CCA TCG ACG ATC CGC GAT TGT CGA TCA 615 Val Pro Lys lie Ala Thr Leu Pro Ser Thr lie Arg Asp Cys Arg Ser 45 50 55 TTA AAG TCT GCC TGC AAC TTA ATT GCT TTA ATT TTA ATA CTG TTA GTC 663 Leu Lys Ser Ala Cys Asn Leu lie Ala Leu lie Leu lie Leu Leu Val 60 65 70 CAT AAG ATA TCC GCA GCT GGT AAC TTC GAG CTG GAA ATA TTA GAA ATC 711 His Lys lie Ser Ala Ala Gly Asn Phe Glu Leu Glu lie Leu Glu lie 75 80 85 90 TCA AAT ACC AAC AGC CAT CTA CTC AAC GGC TAT TGC TGC GGC ATG CCA 759 Ser Asn Thr Asn Ser His Leu Leu Asn Gly Tyr Cys Cys Gly Met Pro 95 100 105 GCG GAA CTT AGG GCC ACC AAG ACG ATA GGC TGC TCG CCA TGC ACG ACG 807 Ala Glu Leu Arg Ala Thr Lys Thr lie Gly Cys Ser Pro Cys Thr Thr 110 115 120 GCA TTC CGG CTG TGC CTG AAG GAG TAC CAG ACC ACG GAG CAG GGT GCC 855 Ala Phe Arg Leu Cys Leu Lys Glu Tyr Gin Thr Thr Glu Gin Gly Ala 125 130 135 AGC ATA TCC ACG GGC TGT TCG TTT GGC AAC GCC ACC ACC AAG ATA CTG 903 Ser lie Ser Thr Gly Cys Ser Phe Gly Asn Ala Thr Thr Lys lie Leu 140 145 150 GGT GGC TCC AGC TTT GTG CTC AGC GAT CCG GGT GTG GGA GCC ATT GTG 951 Gly Gly Ser Ser Phe Val Leu Ser Asp Pro Gly Val Gly Ala lie Val 155 160 165 170 CTG CCC TTT ACG TTT CGT TGG ACG AAG TCG TTT ACG CTG ATA CTG CAG 999 Leu Pro Phe Thr Phe Arg Trp Thr Lys Ser Phe Thr Leu lie Leu Gin 175 180 185 GCG TTG GAT ATG TAC AAC ACA TCC TAT CCA GAT GCG GAG AGG TTA ATT 1047 -97- Ala Leu Asp Met Tyr Asn Thr Ser Tyr Pro Asp Ala Glu Arg Leu lie 190 195 200 GAG GAA ACA TCA TAC TCG GGC GTG ATA CTG CCG TCG CCG GAG TGG AAG 1095 Glu Glu Thr Ser Tyr Ser Gly Val lie Leu Pro Ser Pro Glu Trp Lys 205 210 215 ACG CTG GAC CAC ATC GGG CGG AAC GCG CGG ATC ACC TAC CGT GTC CGG 1143 Thr Leu Asp His lie Gly Arg Asn Ala Arg lie Thr Tyr Arg Val Arg 220 225 230 GTG CAA TGC GCC GTT ACC TAC TAC AAC ACG ACC TGC ACG ACC TTC TGC 1191 Val Gin Cys Ala Val Thr Tyr Tyr Asn Thr Thr Cys Thr Thr Phe Cys 235 240 245 250 CGT CCG CGG GAC GAT CAG TTC GGT CAC TAC GCC TGC GGC TCC GAG GGT 1239 Arg Pro Arg Asp Asp Gin Phe Gly His Tyr Ala Cys Gly Ser Glu Gly 255 260 265 CAG AAG CTC TGC CTG AAT GGC TGG CAG GGC GTC AAC TGC GAG GAG GCC 1287 Gin Lys Leu Cys Leu Asn Gly Trp Gin Gly Val Asn Cys Glu Glu Ala 270 275 280 ATA TGC AAG GCG GGC TGC GAC CCC GTC CAC GGC 1320 lie Cys Lys Ala Gly Cys Asp Pro Val His Gly 285 290 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 293 amino acids (B) TYPE: amino acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: Met Phe Arg Lys His Phe Arg Arg Lys Pro Ala Thr Ser Ser Ser Leu 1 5 10 15 Glu Ser Thr lie Glu Ser Ala Asp Ser Leu Gly Met Ser Lys Lys Thr 20 25 30 Ala Thr Lys Arg Gin Arg Pro Arg His Arg Val Pro Lys lie Ala Thr 35 40 45 Leu Pro Ser Thr lie Arg Asp Cys Arg Ser Leu Lys Ser Ala Cys Asn 50 55 60 Leu lie Ala Leu lie Leu lie Leu Leu Val His Lys lie Ser Ala Ala 65 70 75 80 Gly Asn Phe Glu Leu Glu lie Leu Glu lie Ser Asn Thr Asn Ser His 85 90 95 Leu Leu Asn Gly Tyr Cys Cys Gly Met Pro Ala Glu Leu Arg Ala Thr 100 105 110 Lys Thr lie Gly Cys Ser Pro Cys Thr Thr Ala Phe Arg Leu Cys Leu 115 120 125 Lys Glu Tyr Gin Thr Thr Glu Gin Gly Ala Ser lie Ser Thr Gly Cys 130 135 140 Ser Phe Gly Asn Ala Thr Thr Lys lie Leu Gly Gly Ser Ser Phe Val -98- 145 150 155 160 Leu Ser Asp Pro Gly Val Gly Ala lie Val Leu Pro Phe Thr Phe Arg 165 170 175 Trp Thr Lys Ser Phe Thr Leu lie Leu Gin Ala Leu Asp Met Tyr Asn 180 185 190 Thr Ser Tyr Pro Asp Ala Glu Arg Leu He Glu Glu Thr Ser Tyr Ser 195 200 205 Gly Val He Leu Pro Ser Pro Glu Trp Lys Thr Leu Asp His He Gly 210 215 220 Arg Asn Ala Arg He Thr Tyr Arg Val Arg Val Gin Cys Ala Val Thr 225 230 235 240 Tyr Tyr Asn Thr Thr Cys Thr Thr Phe Cys Arg Pro Arg Asp Asp Gin 245 250 255 Phe Gly His Tyr Ala Cys Gly Ser Glu Gly Gin Lys Leu Cys Leu Asn 260 265 270 Gly Trp Gin Gly Val Asn Cys Glu Glu Ala He Cys Lys Ala Gly Cys 275 280 285 Asp Pro Val His Gly 290 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 267 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: CGGTGGACTT CCTTCGTGTA TTGGTGGGAG CCCTCGGGAA CGGGGGGTAA CACTGAAAGG 60 TCGAGTACCC ATTTCCGTCA TAACGGGTTG GTCGCCCCCT AGGGGTCGGA GTCAGGTGGA 120 CGGGAGGTCG ACAACGCCCG GGGGACGGGT GGTACATGGT GTAAGGTCTT TACCGGACCG 180 GGCAAACGGG TCACACCGAA AGGGGTGAAC GGTAACTACG GGGTCGTCCT GCCCGTCCAT 240 CGAGTCTGGT AAGAGGGTCG CCTTAAG 267 (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 574 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: cDNA -99- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: GAATTCCTTC CATTATACGT GACTTTTCTG AAACTGTAGC CACCCTAGTG TCTCTAACTC 60 CCTCTGGAGT TTGTCAGCTT TGGTCTTTTC AAAGAGCAGG CTCTCTTCAA GCTCCTTAAT 120 GCGGGCATGC TCCAGTTTGG TCTGCGTCTC AAGATCACCT TTGGTAATTG ATTCTTCTTC 180 AACCCGGAAC TGAAGGCTGG CTCTCACCCT CTAGGCAGAG CAGGAATTCC GAGGTGGATG 240 TGTTAGATGT GAATGTCCGT GGCCCAGATG GCTGCACCCC ATTGATGTTG GCTTCTCTCC 300 GAGGAGGCAG CTCAGATTTG AGTGATGAAG ATGAAGATGC AGAGGACTGT TCTGCTAACA 360 TCATCACAGA CTTGGTCTAC CAGGGTGCCA GCCTCCAGNC CAGACAGACC GGACTGGTGA 420 GATGGCCCTG CACCTTGCAG CCCGCTACTC ACGGGCTGAT GCTGCCAAGC GTCTCCTGGA 480 TGCAGGTGCA GATGCCAATG CCCAGGACAA CATGGGCCGC TGTCCACTCC ATGCTGCAGT 540 GGCACGTGAT GCCAAGGTGT ATTCAGATCT GTTA 574 (2) INFORMATION FOR SEQ ID NO:7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 295 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: TCCAGATTCT GATTCGCAAC CGAGTAACTG ATCTAGATGC CAGGATGAAT GATGGTACTA 60 CACCCCTGAT CCTGGCTGCC CGCCTGGCTG TGGAGGGAAT GGTGGCAGAA CTGATCAACT 120 GCCAAGCGGA TGTGAATGCA GTGGATGACC ATGGAAAATC TGCTCTTCAC TGGGCAGCTG 180 CTGTCAATAA TGTGGAGGCA ACTCTTTTGT TGTTGAAAAA TGGGGCCAAC CGAGACATGC 240 AGGACAACAA GGAAGAGACA CCTCTGTTTC TTGCTGCCCG GGAGGAGCTA TAAGC 295 (2) INFORMATION FOR SEQ ID NO:8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 248 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: GAATTCCATT CAGGAGGAAA GGGTGGGGAG AGAAGCAGGC ACCCACTTTC CCGTGGCTGG 60 ACTCGTTCCC AGGTGGCTCC ACCGGCAGCT GTGACCGCCG CAGGTGGGGG CGGAGTGCCA 120 TTCAGAAAAT TCCAGAAAAG CCCTACCCCA ACTCGGACGG CAACGTCACA CCCGTGGGTA 180 -100- GCAACTGGCA CACAAACAGC CAGCGTGTCT GGGGCACGGG GGGATGGCAC CCCCTGCAGG 240 CAGAGCTG 248 (2) INFORMATION FOR SEQ ID NO:9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 323 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: TACGTATCTC GAGCACAGAC AGCTGACGTA CACTTTTNNA GTGCGAGGGA CATTCGTCCG 60 ACCAGTACGA ACATTTAGGC TCAGTACGGT AGGTCCATGG CCAAGACTAG GAGACGTAGG 120 GAGCTACAGG TCCCGCTCGC TAAACTCGGA CCACTGAAAC CTCCGGTCGA CAGTCGGTAA 180 GCGAACAAGA GGGCCAGATC TTAGAGAAGG TGTCGCGGCG AGACTCGGGC TCGGGTCAGG 240 CGGCCTTAAG GACGTCGGGC CCNNNAGGTG ATCAAGATCT CGNCNCGGCG GGCGCCACCT 300 CGAGGNCGAA AACAAGGGAA ATC 323 (2) INFORMATION FOR SEQ ID NO:10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3234 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: cDNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..3234 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10: TGC CAG GAG GAC GCG GGC AAC AAG GTC TGC AGC CTG CAG TGC AAC AAC 48 Cys Gin Glu Asp Ala Gly Asn Lys Val Cys Ser Leu Gin Cys Asn Asn 1 5 10 15 CAC GCG TGC GGC TGG GAC GGC GGT GAC TGC TCC CTC AAC TTC AAT GAC 96 His Ala Cys Gly Trp Asp Gly Gly Asp Cys Ser Leu Asn Phe Asn Asp 20 25 30 CCC TGG AAG AAC TGC ACG CAG TCT CTG CAG TGC TGG AAG TAC TTC AGT 144 Pro Trp Lys Asn Cys Thr Gin Ser Leu Gin Cys Trp Lys Tyr Phe Ser 35 40 45 GAC GGC CAC TGT GAC AGC CAG TGC AAC TCA GCC GGC TGC CTC TTC GAC 192 Asp Gly His Cys Asp Ser Gin Cys Asn Ser Ala Gly Cys Leu Phe Asp 50 55 60 GGC TTT GAC TGC CAG CGT GCG GAA GGC CAG TGC AAC CCC CTG TAC GAC 240 -101- Gly Phe Asp Cys Gin Arg Ala Glu Gly Gin Cys Asn Pro Leu Tyr Asp 65 70 75 80 CAG TAC TGC AAG GAC CAC TTC AGC GAC GGG CAC TGC GAC CAG GGC TGC 288 Gin Tyr Cys Lys Asp His Phe Ser Asp Gly His Cys Asp Gin Gly Cys 85 90 95 AAC AGC GCG GAG TGC GAG TGG GAC GGG CTG GAC TGT GCG GAG CAT GTA 336 Asn Ser Ala Glu Cys Glu Trp Asp Gly Leu Asp Cys Ala Glu His Val 100 105 110 CCC GAG AGG CTG GCG GCC GGC ACG CTG GTG GTG GTG GTG CTG ATG CCG 384 Pro Glu Arg Leu Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro 115 120 125 CCG GAG CAG CTG CGC AAC AGC TCC TTC CAC TTC CTG CGG GAG CTC AGC 432 Pro Glu Gin Leu Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser 130 135 140 CGC GTG CTG CAC ACC AAC GTG GTC TTC AAG CGT GAC GCA CAC GGC CAG 480 Arg Val Leu His Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gin 145 150 155 160 CAG ATG ATC TTC CCC TAC TAC GGC CGC GAG GAG GAG CTG CGC AAG CAC 528 Gin Met He Phe Pro Tyr Tyr Gly Arg Glu Glu Glu Leu Arg Lys His 165 170 175 CCC ATC AAG CGT GCC GCC GAG GGC TGG GCC GCA CCT GAC GCC CTG CTG 576 Pro He Lys Arg Ala Ala Glu Gly Trp Ala Ala Pro Asp Ala Leu Leu 180 185 190 GGC CAG GTG AAG GCC TCG CTG CTC CCT GGT GGC AGC GAG GGT GGG CGG 624 Gly Gin Val Lys Ala Ser Leu Leu Pro Gly Gly Ser Glu Gly Gly Arg 195 200 205 CGG CGG AGG GAG CTG GAC CCC ATG GAC GTC CGC GGC TCC ATC GTC TAC 672 Arg Arg Arg Glu Leu Asp Pro Met Asp Val Arg Gly Ser He Val Tyr 210 215 220 CTG GAG ATT GAC AAC CGG CAG TGT GTG CAG GCC TCC TCG CAG TGC TTC 720 Leu Glu He Asp Asn Arg Gin Cys Val Gin Ala Ser Ser Gin Cys Phe 225 230 235 240 CAG AGT GCC ACC GAC GTG GCC GCA TTC CTG GGA GCG CTC GCC TCG CTG 768 Gin Ser Ala Thr Asp Val Ala Ala Phe Leu Gly Ala Leu Ala Ser Leu 245 250 255 GGC AGC CTC AAC ATC CCC TAC AAG ATC GAG GCC GTG CAG AGT GAG ACC 816 Gly Ser Leu Asn He Pro Tyr Lys He Glu Ala Val Gin Ser Glu Thr 260 265 270 GTG GAG CCG CCC CCG CCG GCG CAG CTG CAC TTC ATG TAC GTG GCG GCG 864 Val Glu Pro Pro Pro Pro Ala Gin Leu His Phe Met Tyr Val Ala Ala 275 280 285 GCC GCC TTT GTG CTT CTG TTC TTC GTG GGC TGC GGG GTG CTG CTG TCC 912 Ala Ala Phe Val Leu Leu Phe Phe Val Gly Cys Gly Val Leu Leu Ser 290 295 300 CGC AAG CGC CGG CGG CAG CAT GGC CAG CTC TGG TTC CCT GAG GGC TTC 960 Arg Lys Arg Arg Arg Gin His Gly Gin Leu Trp Phe Pro Glu Gly Phe 305 310 315 320 AAA GTG TCT GAG GCC AGC AAG AAG AAG CGG CGG GAG CCC CTC GGC GAG 1008 Lys Val Ser Glu Ala Ser Lys Lys Lys Arg Arg Glu Pro Leu Gly Glu 325 330 335 -102- GAC TCC GTG GGC CTC AAG CCC CTG AAG AAC GCT TCA GAC GGT GCC CTC 1056 Asp Ser Val Gly Leu Lys Pro Leu Lys Asn Ala Ser Asp Gly Ala Leu 340 345 350 ATG GAC GAC AAC CAG AAT GAG TGG GGG GAC GAG GAC CTG GAG ACC AAG 1104 Met Asp Asp Asn Gin Asn Glu Trp Gly Asp Glu Asp Leu Glu Thr Lys 355 360 365 AAG TTC CGG TTC GAG GAG CCC GTG GTT CTG CCT GAC CTG GAC GAC CAG 1152 Lys Phe Arg Phe Glu Glu Pro Val Val Leu Pro Asp Leu Asp Asp Gin 370 375 380 ACA GAC CAC CGG CAG TGG ACT CAG CAG CAC CTG GAT GCC GCT GAC CTG 1200 Thr Asp His Arg Gin Trp Thr Gin Gin His Leu Asp Ala Ala Asp Leu 385 390 395 400 CGC ATG TCT GCC ATG GCC CCC ACA CCG CCC CAG GGT GAG GTT GAC GCC 1248 Arg Met Ser Ala Met Ala Pro Thr Pro Pro Gin Gly Glu Val Asp Ala 405 410 415 GAC TGC ATG GAC GTC AAT GTC CGC GGG CCT GAT GGC TTC ACC CCG CTC 1296 Asp Cys Met Asp Val Asn Val Arg Gly Pro Asp Gly Phe Thr Pro Leu 420 425 430 ATG ATC GCC TCC TGC AGC GGG GGC GGC CTG GAG ACG GGC AAC AGC GAG 1344 Met He Ala Ser Cys Ser Gly Gly Gly Leu Glu Thr Gly Asn Ser Glu 435 440 445 GAA GAG GAG GAC GCG CCG GCC GTC ATC TCC GAC TTC ATC TAC CAG GGC 1392 Glu Glu Glu Asp Ala Pro Ala Val He Ser Asp Phe He Tyr Gin Gly 450 455 460 GCC AGC CTG CAC AAC CAG ACA GAC CGC ACG GGC GAG ACC GCC TTG CAC 1440 Ala Ser Leu His Asn Gin Thr Asp Arg Thr Gly Glu Thr Ala Leu His 465 470 475 480 CTG GCC GCC CGC TAC TCA CGC TCT GAT GCC GCC AAG CGC CTG CTG GAG 1488 Leu Ala Ala Arg Tyr Ser Arg Ser Asp Ala Ala Lys Arg Leu Leu Glu 485 490 495 GCC AGC GCA GAT GCC AAC ATC CAG GAC AAC ATG GGC CGC ACC CCG CTG 1536 Ala Ser Ala Asp Ala Asn He Gin Asp Asn Met Gly Arg Thr Pro Leu 500 505 510 CAT GCG GCT GTG TCT GCC GAC GCA CAA GGT GTC TTC CAG ATC CTG ATC 1584 His Ala Ala Val Ser Ala Asp Ala Gin Gly Val Phe Gin He Leu He 515 520 525 CGG AAC CGA GCC ACA GAC CTG GAT GCC CGC ATG CAT GAT GGC ACG ACG 1632 Arg Asn Arg Ala Thr Asp Leu Asp Ala Arg Met His Asp Gly Thr Thr 530 535 540 CCA CTG ATC CTG GCT GCC CGC CTG GCC GTG GAG GGC ATG CTG GAG GAC 1680 Pro Leu He Leu Ala Ala Arg Leu Ala Val Glu Gly Met Leu Glu Asp 545 550 555 560. CTC ATC AAC TCA CAC GCC GAC GTC AAC GCC GTA GAT GAC CTG GGC AAG 1728 Leu He Asn Ser His Ala Asp Val Asn Ala Val Asp Asp Leu Gly Lys 565 570 575 TCC GCC CTG CAC TGG GCC GCC GCC GTG AAC AAT GTG GAT GCC GCA GTT 1776 Ser Ala Leu His Trp Ala Ala Ala Val Asn Asn Val Asp Ala Ala Val 580 585 590 GTG CTC CTG AAG AAC GGG GCT AAC AAA GAT ATG CAG AAC AAC AGG GAG 1824 Val Leu Leu Lys Asn Gly Ala Asn Lys Asp Met Gin Asn Asn Arg Glu 595 600 605 -103- GAG ACA CCC CTG TTT CTG GCC GCC CGG GAG GGC AGC TAC GAG ACC GCC 1872 Glu Thr Pro Leu Phe Leu Ala Ala Arg Glu Gly Ser Tyr Glu Thr Ala 610 615 620 AAG GTG CTG CTG GAC CAC TTT GCC AAC CGG GAC ATC ACG GAT CAT ATG 1920 Lys Val Leu Leu Asp His Phe Ala Asn Arg Asp He Thr Asp His Met 625 630 635 640 GAC CGC CTG CCG CGC GAC ATC GCA CAG GAG CGC ATG CAT CAC GAC ATC 1968 Asp Arg Leu Pro Arg Asp He Ala Gin Glu Arg Met His His Asp He 645 650 655 GTG AGG CTG CTG GAC GAG TAC AAC CTG GTG CGC AGC CCG CAG CTG CAC 2016 Val Arg Leu Leu Asp Glu Tyr Asn Leu Val Arg Ser Pro Gin Leu His 660 665 670 GGA GCC CCG CTG GGG GGC ACG CCC ACC CTG TCG CCC CCG CTC TGC TCG 2064 Gly Ala Pro Leu Gly Gly Thr Pro Thr Leu Ser Pro Pro Leu Cys Ser 675 680 685 CCC AAC GGC TAC CTG GGC AGC CTC AAG CCC GGC GTG CAG GGC AAG AAG 2112 Pro Asn Gly Tyr Leu Gly Ser Leu Lys Pro Gly Val Gin Gly Lys Lys 690 695 700 GTC CGC AAG CCC AGC AGC AAA GGC CTG GCC TGT GGA AGC AAG GAG GCC 2160 Val Arg Lys Pro Ser Ser Lys Gly Leu Ala Cys Gly Ser Lys Glu Ala 705 710 715 720 AAG GAC CTC AAG GCA CGG AGG AAG AAG TCC CAG GAT GGC AAG GGC TGC 2208 Lys Asp Leu Lys Ala Arg Arg Lys Lys Ser Gin Asp Gly Lys Gly Cys 725 730 735 CTG CTG GAC AGC TCC GGC ATG CTC TCG CCC GTG GAC TCC CTG GAG TCA 2256 Leu Leu Asp Ser Ser Gly Met Leu Ser Pro Val Asp Ser Leu Glu Ser 740 745 750 CCC CAT GGC TAC CTG TCA GAC GTG GCC TCG CCG CCA CTG CTG CCC TCC 2304 Pro His Gly Tyr Leu Ser\Asp Val Ala Ser Pro Pro Leu Leu Pro Ser 755 760 765 CCG TTC CAG CAG TCT CCG TCC GTG CCC CTC AAC CAC CTG CCT GGG ATG 2352 Pro Phe Gin Gin Ser Pro Ser Val Pro Leu Asn His Leu Pro Gly Met 770 775 780 CCC GAC ACC CAC CTG GGC ATC GGG CAC CTG AAC GTG GCG GCC AAG CCC 2400 Pro Asp Thr His Leu Gly He Gly His Leu Asn Val Ala Ala Lys Pro 785 790 795 800 GAG ATG GCG GCG CTG GGT GGG GGC GGC CGG CTG GCC TTT GAG ACT GGC 2448 Glu Met Ala Ala Leu Gly Gly Gly Gly Arg Leu Ala Phe Glu Thr Gly 805 810 815 CCA CCT CGT CTC TCC CAC CTG CCT GTG GCC TCT GGC ACC AGC ACC GTC 2496 Pro Pro Arg Leu Ser His Leu Pro Val Ala Ser Gly Thr Ser Thr Val 820 825 830 CTG GGC TCC AGC AGC GGA GGG GCC CTG AAT TTC ACT GTG GGC GGG TCC 2544 Leu Gly Ser Ser Ser Gly Gly Ala Leu Asn Phe Thr Val Gly Gly Ser 835 840 845 ACC AGT TTG AAT GGT CAA TGC GAG TGG CTG TCC CGG CTG CAG AGC GGC 2592 Thr Ser Leu Asn Gly Gin Cys Glu Trp Leu Ser Arg Leu Gin Ser Gly 850 855 860 ATG GTG CCG AAC CAA TAC AAC CCT CTG CGG GGG AGT GTG GCA CCA GGC 2640 Met Val Pro Asn Gin Tyr Asn Pro Leu Arg Gly Ser Val Ala Pro Gly 865 870 875 880 -104- CCC CTG AGC ACA CAG GCC CCC TCC CTG CAG CAT GGC ATG GTA GGC CCG 2688 Pro Leu Ser Thr Gin Ala Pro Ser Leu Gin His Gly Met Val Gly Pro 885 890 895 CTG CAC AGT AGC CTT GCT GCC AGC GCC CTG TCC CAG ATG ATG AGC TAC 2736 Leu His Ser Ser Leu Ala Ala Ser Ala Leu Ser Gin Met Met Ser Tyr 900 905 910 CAG GGC CTG CCC AGC ACC CGG CTG GCC ACC CAG CCT CAC CTG GTG CAG 2784 Gin Gly Leu Pro Ser Thr Arg Leu Ala Thr Gin Pro His Leu Val Gin 915 920 925 ACC CAG CAG GTG CAG CCA CAA AAC TTA CAG ATG CAG CAG CAG AAC CTG 2832 Thr Gin Gin Val Gin Pro Gin Asn Leu Gin Met Gin Gin Gin Asn Leu 930 935 940 CAG CCA GCA AAC ATC CAG CAG CAG CAA AGC CTG CAG CCG CCA CCA CCA 2880 Gin Pro Ala Asn He Gin Gin Gin Gin Ser Leu Gin Pro Pro Pro Pro 945 950 955 960 CCA CCA CAG CCG CAC CTT GGC GTG AGC TCA GCA GCC AGC GGC CAC CTG 2928 Pro Pro Gin Pro His Leu Gly Val Ser Ser Ala Ala Ser Gly His Leu 965 970 975 GGC CGG AGC TTC CTG AGT GGA GAG CCG AGC CAG GCA GAC GTG CAG CCA 2976 Gly Arg Ser Phe Leu Ser Gly Glu Pro Ser Gin Ala Asp Val Gin Pro 980 985 990 CTG GGC CCC AGC AGC CTG GCG GTG CAC ACT ATT CTG CCC CAG GAG AGC 3024 Leu Gly Pro Ser Ser Leu Ala Val His Thr He Leu Pro Gin Glu Ser 995 1000 1005 CCC GCC CTG CCC ACG TCG CTG CCA TCC TCG CTG GTC CCA CCC GTG ACC 3072 Pro Ala Leu Pro Thr Ser Leu Pro Ser Ser Leu Val Pro Pro Val Thr 1010 1015 1020 GCA GCC CAG TTC CTG ACG CCC CCC TCG CAG CAC AGC TAC TCC TCG CCT 3120 Ala Ala Gin Phe Leu Thr Pro Pro Ser Gin His Ser Tyr Ser Ser Pro 1025 1030 1035 1040 GTG GAC AAC ACC CCC AGC CAC CAG CTA CAG GTG CCT GTT CCT GTA ATG 3168 Val Asp Asn Thr Pro Ser His Gin Leu Gin Val Pro Val Pro Val Met 1045 1050 1055 GTA ATG ATC CGA TCT TCG GAT CCT TCT AAA GGC TCA TCA ATT TTG ATC 3216 Val Met He Arg Ser Ser Asp Pro Ser Lys Gly Ser Ser He Leu He 1060 1065 1070 GAA GCT CCC GAC TCA TGG 3234 Glu Ala Pro Asp Ser Trp 1075 (2) INFORMATION FOR SEQ ID NO:11: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1078 amino acids (B) TYPE: amino acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: Cys Gin Glu Asp Ala Gly Asn Lys Val Cys Ser Leu Gin Cys Asn Asn 1 5 10 15 -105- His Ala Cys Gly Trp Asp Gly Gly Asp Cys Ser Leu Asn Phe Asn Asp 20 25 30 Pro Trp Lys Asn Cys Thr Gin Ser Leu Gin Cys Trp Lys Tyr Phe Ser 35 40 45 Asp Gly His Cys Asp Ser Gin Cys Asn Ser Ala Gly Cys Leu Phe Asp 50 55 60 Gly Phe Asp Cys Gin Arg Ala Glu Gly Gin Cys Asn Pro Leu Tyr Asp 65 70 75 80 Gin Tyr Cys Lys Asp His Phe Ser Asp Gly His Cys Asp Gin Gly Cys 85 90 95 Asn Ser Ala Glu Cys Glu Trp Asp Gly Leu Asp Cys Ala Glu His Val 100 105 110 Pro Glu Arg Leu Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro 115 120 125 Pro Glu Gin Leu Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser 130 135 140 Arg Val Leu His Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gin 145 150 155 160 Gin Met He Phe Pro Tyr Tyr Gly Arg Glu Glu Glu Leu Arg Lys His 165 170 175 Pro He Lys Arg Ala Ala Glu Gly Trp Ala Ala Pro Asp Ala Leu Leu 180 185 190 Gly Gin Val Lys Ala Ser Leu Leu Pro Gly Gly Ser Glu Gly Gly Arg 195 200 205 Arg Arg Arg Glu Leu Asp Pro Met Asp Val Arg Gly Ser He Val Tyr 210 215 220 Leu Glu He Asp Asn Arg Gin Cys Val Gin Ala Ser Ser Gin Cys Phe 225 230 235 240 Gin Ser Ala Thr Asp Val Ala Ala Phe Leu Gly Ala Leu Ala Ser Leu 245 250 255 Gly Ser Leu Asn He Pro Tyr Lys He Glu Ala Val Gin Ser Glu Thr 260 265 270 Val Glu Pro Pro Pro Pro Ala Gin Leu His Phe Met Tyr Val Ala Ala 275 280 285 Ala Ala Phe Val Leu Leu Phe Phe Val Gly Cys Gly Val Leu Leu Ser 290 295 300 Arg Lys Arg Arg Arg Gin His Gly Gin Leu Trp Phe Pro Glu Gly Phe 305 310 315 320 Lys Val Ser Glu Ala Ser Lys Lys Lys Arg Arg Glu Pro Leu Gly Glu 325 330 335 Asp Ser Val Gly Leu Lys Pro Leu Lys Asn Ala Ser Asp Gly Ala Leu 340 345 350 Met Asp Asp Asn Gin Asn Glu Trp Gly Asp Glu Asp Leu Glu Thr Lys 355 360 365 Lys Phe Arg Phe Glu Glu Pro Val Val Leu Pro Asp Leu Asp Asp Gin -106- 370 375 380 Thr Asp His Arg Gin Trp Thr Gin Gin His Leu Asp Ala Ala Asp Leu 385 390 395 400 Arg Met Ser Ala Met Ala Pro Thr Pro Pro Gin Gly Glu Val Asp Ala 405 410 415 Asp Cys Met Asp Val Asn Val Arg Gly Pro Asp Gly Phe Thr Pro Leu 420 425 430 Met He Ala Ser Cys Ser Gly Gly Gly Leu Glu Thr Gly Asn Ser Glu 435 440 445 Glu Glu Glu Asp Ala Pro Ala Val He Ser Asp Phe He Tyr Gin Gly 450 455 460 Ala Ser Leu His Asn Gin Thr Asp Arg Thr Gly Glu Thr Ala Leu His 465 470 475 480 Leu Ala Ala Arg Tyr Ser Arg Ser Asp Ala Ala Lys Arg Leu Leu Glu 485 490 495 Ala Ser Ala Asp Ala Asn He Gin Asp Asn Met Gly Arg Thr Pro Leu 500 505 510 His Ala Ala Val Ser Ala Asp Ala Gin Gly Val Phe Gin He Leu He 515 520 525 Arg Asn Arg Ala Thr Asp Leu Asp Ala Arg Met His Asp Gly Thr Thr 530 535 540 Pro Leu He Leu Ala Ala Arg Leu Ala Val Glu Gly Met Leu Glu Asp 545 550 555 560 Leu He Asn Ser His Ala Asp Val Asn Ala Val Asp Asp Leu Gly Lys 565 570 575 Ser Ala Leu His Trp Ala Ala Ala Val Asn Asn Val Asp Ala Ala Val 580 585 590 Val Leu Leu Lys Asn Gly Ala Asn Lys Asp Met Gin Asn Asn Arg Glu 595 600 605 Glu Thr Pro Leu Phe Leu Ala Ala Arg Glu Gly Ser Tyr Glu Thr Ala 610 615 620 Lys Val Leu Leu Asp His Phe Ala Asn Arg Asp He Thr Asp His Met 625 630 635 640 Asp Arg Leu Pro Arg Asp He Ala Gin Glu Arg Met His His Asp He 645 650 655 Val Arg Leu Leu Asp Glu Tyr Asn Leu Val Arg Ser Pro Gin Leu His 660 665 670 Gly Ala Pro Leu Gly Gly Thr Pro Thr Leu Ser Pro Pro Leu Cys Ser 675 680 685 Pro Asn Gly Tyr Leu Gly Ser Leu Lys Pro Gly Val Gin Gly Lys Lys 690 695 700 Val Arg Lys Pro Ser Ser Lys Gly Leu Ala Cys Gly Ser Lys Glu Ala 705 710 715 720 Lys Asp Leu Lys Ala Arg Arg Lys Lys Ser Gin Asp Gly Lys Gly Cys 725 730 735 -107- Leu Leu Asp Ser Ser Gly Met Leu Ser Pro Val Asp Ser Leu Glu Ser 740 745 750 Pro His Gly Tyr Leu Ser Asp Val Ala Ser Pro Pro Leu Leu Pro Ser 755 760 765 Pro Phe Gin Gin Ser Pro Ser Val Pro Leu Asn His Leu Pro Gly Met 770 775 780 Pro Asp Thr His Leu Gly He Gly His Leu Asn Val Ala Ala Lys Pro 785 790 795 800 Glu Met Ala Ala Leu Gly Gly Gly Gly Arg Leu Ala Phe Glu Thr Gly 805 810 815 Pro Pro Arg Leu Ser His Leu Pro Val Ala Ser Gly Thr Ser Thr Val 820 825 830 Leu Gly Ser Ser Ser Gly Gly Ala Leu Asn Phe Thr Val Gly Gly Ser 835 840 845 Thr Ser Leu Asn Gly Gin Cys Glu Trp Leu Ser Arg Leu Gin Ser Gly 850 855 860 Met Val Pro Asn Gin Tyr Asn Pro Leu Arg Gly Ser Val Ala Pro Gly 865 870 875 880 Pro Leu Ser Thr Gin Ala Pro Ser Leu Gin His Gly Met Val Gly Pro 885 890 895 Leu His Ser Ser Leu Ala Ala Ser Ala Leu Ser Gin Met Met Ser Tyr 900 905 910 Gin Gly Leu Pro Ser Thr Arg Leu Ala Thr Gin Pro His Leu Val Gin 915 920 925 Thr Gin Gin Val Gin Pro Gin Asn Leu Gin Met Gin Gin Gin Asn Leu 930 935 940 Gin Pro Ala Asn He Gin Gin Gin Gin Ser Leu Gin Pro Pro Pro Pro 945 950 955 960 Pro Pro Gin Pro His Leu Gly Val Ser Ser Ala Ala Ser Gly His Leu 965 970 975 Gly Arg Ser Phe Leu Ser Gly Glu Pro Ser Gin Ala Asp Val Gin Pro 980 985 990 Leu Gly Pro Ser Ser Leu Ala Val His Thr He Leu Pro Gin Glu Ser 995 1000 1005 Pro Ala Leu Pro Thr Ser Leu Pro Ser Ser Leu Val Pro Pro Val Thr 1010 1015 1020 Ala Ala Gin Phe Leu Thr Pro Pro Ser Gin His Ser Tyr Ser Ser Pro 1025 1030 1035 1040 Val Asp Asn Thr Pro Ser His Gin Leu Gin Val Pro Val Pro Val Met 1045 1050 1055 Val Met He Arg Ser Ser Asp Pro Ser Lys Gly Ser Ser He Leu He 1060 1065 1070 Glu Ala Pro Asp Ser Trp 1075 (2) INFORMATION FOR SEQ ID NO:12: -108- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4268 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: cDNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 2..1972 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: G GAG GTG GAT GTG TTA GAT GTG AAT GTC CGT GGC CCA GAT GGC TGC 46 Glu Val Asp Val Leu Asp Val Asn Val Arg Gly Pro Asp Gly Cys 1 5 10 15 ACC CCA TTG ATG TTG GCT TCT CTC CGA GGA GGC AGC TCA GAT TTG AGT 94 Thr Pro Leu Met Leu Ala Ser Leu krg Gly Gly Ser Ser Asp Leu Ser 20 25 30 GAT GAA GAT GAA GAT GCA GAG GAC TCT TCT GCT AAC ATC ATC ACA GAC 142 Asp Glu Asp Glu Asp Ala Glu Asp Ser Ser Ala Asn He He Thr Asp 35 40 45 TTG GTC TAC CAG GGT GCC AGC CTC CAG GCC CAG ACA GAC CGG ACT GGT 190 Leu Val Tyr Gin Gly Ala Ser Leu Gin Ala Gin Thr Asp Arg Thr Gly 50 55 60 GAG ATG GCC CTG CAC CTT GCA GCC CGC TAC TCA CGG GCT GAT GCT GCC 238 Glu Met Ala Leu His Leu Ala Ala Arg Tyr Ser Arg Ala Asp Ala Ala 65 70 75 AAG CGT CTC CTG GAT GCA GGT GCA GAT GCC AAT GCC CAG GAC AAC ATG 286 Lys Arg Leu Leu Asp Ala Gly Ala Asp Ala Asn Ala Gin Asp Asn Met 80 85 90 95 GGC CGC TGT CCA CTC CAT GCT GCA GTG GCA GCT GAT GCC CAA GGT GTC 334 Gly Arg Cys Pro Leu His Ala Ala Val Ala Ala Asp Ala Gin Gly Val 100 105 110 TTC CAG ATT CTG ATT CGC AAC CGA GTA ACT GAT CTA GAT GCC AGG ATG 382 Phe Gin He Leu He Arg Asn Arg Val Thr Asp Leu Asp Ala Arg Met 115 120 125 AAT GAT GGT ACT ACA CCC CTG ATC CTG GCT GCC CGC CTG GCT GTG GAG 430 Asn Asp Gly Thr Thr Pro Leu He Leu Ala Ala Arg Leu Ala Val Glu 130 135 140 GGA ATG GTG GCA GAA CTG ATC AAC TGC CAA GCG GAT GTG AAT GCA GTG 478 Gly Met Val Ala Glu Leu He Asn Cys Gin Ala Asp Val Asn Ala Val 145 150 155 GAT GAC CAT GGA AAA TCT GCT CTT CAC TGG GCA GCT GCT GTC AAT AAT 526 Asp Asp His Gly Lys Ser Ala Leu His Trp Ala Ala Ala Val Asn Asn 160 165 170 175 GTG GAG GCA ACT CTT TTG TTG TTG AAA AAT GGG GCC AAC CGA GAC ATG 574 Val Glu Ala Thr Leu Leu Leu Leu Lys Asn Gly Ala Asn Arg Asp Met 180 185 190 CAG GAC AAC AAG GAA GAG ACA CCT CTG TTT CTT GCT GCC CGG GAG GGG 622 Gin Asp Asn Lys Glu Glu Thr Pro Leu Phe Leu Ala Ala Arg Glu Gly 195 200 205 -109- AGC TAT GAA GCA GCC AAG ATC CTG TTA GAC CAT TTT GCC AAT CGA GAC 670 Ser Tyr Glu Ala Ala Lys He Leu Leu Asp His Phe Ala Asn Arg Asp 210 215 220 ATC ACA GAC CAT ATG GAT CGT CTT CCC CGG GAT GTG GCT CGG GAT CGC 718 He Thr Asp His Met Asp Arg Leu Pro Arg Asp Val Ala Arg Asp Arg 225 230 235 ATG CAC CAT GAC ATT GTG CGC CTT CTG GAT GAA TAC AAT GTG ACC CCA 766 Met His His Asp He Val Arg Leu Leu Asp Glu Tyr Asn Val Thr Pro 240 245 250 255 AGC CCT CCA GGC ACC GTG TTG ACT TCT GCT CTC TCA CCT GTC ATC TGT 814 Ser Pro Pro Gly Thr Val Leu Thr Ser Ala Leu Ser Pro Val He Cys 260 265 270 GGG CCC AAC AGA TCT TTC CTC AGC CTG AAG CAC ACC CCA ATG GGC AAG 862 Gly Pro Asn Arg Ser Phe Leu Ser Leu Lys His Thr Pro Met Gly Lys 275 280 285 AAG TCT AGA CGG CCC AGT GCC AAG AGT ACC ATG CCT ACT AGC CTC CCT 910 Lys Ser Arg Arg Pro Ser Ala Lys Ser Thr Met Pro Thr Ser Leu Pro 290 295 300 AAC CTT GCC AAG GAG GCA AAG GAT GCC AAG GGT AGT AGG AGG AAG AAG 958 Asn Leu Ala Lys Glu Ala Lys Asp Ala Lys Gly Ser Arg Arg Lys Lys 305 310 315 TCT CTG AGT GAG AAG GTC CAA CTG TCT GAG AGT TCA GTA ACT TTA TCC 1006 Ser Leu Ser Glu Lys Val Gin Leu Ser Glu Ser Ser Val Thr Leu Ser 320 325 330 335 CCT GTT GAT TCC CTA GAA TCT CCT CAC ACG TAT GTT TCC GAC ACC ACA 1054 Pro Val Asp Ser Leu Glu Ser Pro His Thr Tyr Val Ser Asp Thr Thr 340 345 350 TCC TCT CCA ATG ATT ACA TCC CCT GGG ATC TTA CAG GCC TCA CCC AAC 1102 Ser Ser Pro Met He Thr Ser Pro Gly He Leu Gin Ala Ser Pro Asn 355 360 365 CCT ATG TTG GCC ACT GCC GCC CCT CCT GCC CCA GTC CAT GCC CAG CAT 1150 Pro Met Leu Ala Thr Ala Ala Pro Pro Ala Pro Val His Ala Gin His 370 375 380 GCA CTA TCT TTT TCT AAC CTT CAT GAA ATG CAG CCT TTG GCA CAT GGG 1198 Ala Leu Ser Phe Ser Asn Leu His Glu Met Gin Pro Leu Ala His Gly 385 390 395 GCC AGC ACT GTG CTT CCC TCA GTG AGC CAG TTG CTA TCC CAC CAC CAC 1246 Ala Ser Thr Val Leu Pro Ser Val Ser Gin Leu Leu Ser His His His 400 405 410 415 ATT GTG TCT CCA GGC AGT GGC AGT GCT GGA AGC TTG AGT AGG CTC CAT 1294 He Val Ser Pro Gly Ser Gly Ser Ala Gly Ser Leu Ser Arg Leu His 420 425 430 CCA GTC CCA GTC CCA GCA GAT TGG ATG AAC CGC ATG GAG GTG AAT GAG 1342 Pro Val Pro Val Pro Ala Asp Trp Met Asn Arg Met Glu Val Asn Glu 435 440 445 ACC CAG TAC AAT GAG ATG TTT GGT ATG GTC CTG GCT CCA GCT GAG GGC 1390 Thr Gin Tyr Asn Glu Met Phe Gly Met Val Leu Ala Pro Ala Glu Gly 450 455 460 ACC CAT CCT GGC ATA GCT CCC CAG AGC AGG CCA CCT GAA GGG AAG CAC 1438 Thr His Pro Gly He Ala Pro Gin Ser Arg Pro Pro Glu Gly Lys His 465 470 475 -110- ATA ACC ACC CCT CGG GAG CCC TTG CCC CCC ATT GTG ACT TTC CAG CTC 1486 He Thr Thr Pro Arg Glu Pro Leu Pro Pro He Val Thr Phe Gin Leu 480 485 490 495 ATC CCT AAA GGC AGT ATT GCC CAA CCA GCG GGG GCT CCC CAG CCT CAG 1534 He Pro Lys Gly Ser He Ala Gin Pro Ala Gly Ala Pro Gin Pro Gin 500 505 510 TCC ACC TGC CCT CCA GCT GTT GCG GGC CCC CTG CCC ACC ATG TAC CAG 1582 Ser Thr Cys Pro Pro Ala Val Ala Gly Pro Leu Pro Thr Met Tyr Gin 515 520 525 ATT CCA GAA ATG GCC CGT TTG CCC AGT GTG GCT TTC CCC ACT GCC ATG 1630 He Pro Glu Met Ala Arg Leu Pro Ser Val Ala Phe Pro Thr Ala Met 530 535 540 ATG CCC CAG CAG GAC GGG CAG GTA GCT CAG ACC ATT CTC CCA GCC TAT 1678 Met Pro Gin Gin Asp Gly Gin Val Ala Gin Thr He Leu Pro Ala Tyr 545 550 555 CAT CCT TTC CCA GCC TCT GTG GGC AAG TAC CCC ACA CCC CCT TCA CAG 1726 His Pro Phe Pro Ala Ser Val Gly Lys Tyr Pro Thr Pro Pro Ser Gin 560 565 570 575 CAC AGT TAT GCT TCC TCA AAT GCT GCT GAG CGA ACA CCC AGT CAC AGT 1774 His Ser Tyr Ala Ser Ser Asn Ala Ala Glu Arg Thr Pro Ser His Ser 580 585 590 GGT CAC CTC CAG GGT GAG CAT CCC TAC CTG ACA CCA TCC CCA GAG TCT 1822 Gly His Leu Gin Gly Glu His Pro Tyr Leu Thr Pro Ser Pro Glu Ser 595 600 605 CCT GAC CAG TGG TCA AGT TCA TCA CCC CAC TCT GCT TCT GAC TGG TCA 1870 Pro Asp Gin Trp Ser Ser Ser Ser Pro His Ser Ala Ser Asp Trp Ser 610 615 620 GAT GTG ACC ACC AGC CCT ACC CCT GGG GGT GCT GGA GGA GGT CAG CGG 1918 Asp Val Thr Thr Ser Pro Thr Pro Gly Gly Ala Gly Gly Gly Gin Arg 625 630 635 GGA CCT GGG ACA CAC ATG TCT GAG CCA CCA CAC AAC AAC ATG CAG GTT 1966 Gly Pro Gly Thr His Met Ser Glu Pro Pro His Asn Asn Met Gin Val 640 645 650 655 TAT GCG TGAGAGAGTC CACCTCCAGT GTAGAGACAT AACTGACTTT TGTAAATGCT 2022 Tyr Ala GCTGAGGAAC AAATGAAGGT CATCCGGGAG AGAAATGAAG AAATCTCTGG AGCCAGCTTC 2082 TAGAGGTAGG AAAGAGAAGA TGTTCTTATT CAGATAATGC AAGAGAAGCA ATTCGTCAGT 2142 TTCACTGGGT ATCTGCAAGG CTTATTGATT ATTCTAATCT AATAAGACAA GTTTGTGGAA 2202 ATGCAAGATG AATACAAGCC TTGGGTCCAT GTTTACTCTC TTCTATTTGG AGAATAAGAT 2262 GGATGCTTAT TGAAGCCCAG ACATTCTTGC AGCTTGGACT GCATTTTAAG CCCTGCAGGC 2322 TTCTGCCATA TCCATGAGAA GATTCTACAC TAGCGTCCTG TTGGGAATTA TGCCCTGGAA 2382 TTCTGCCTGA ATTGACCTAC GCATCTCCTC CTCCTTGGAC ATTCTTTTGT CTTCATTTGG 2442 TGCTTTTGGT TTTGCACCTC TCCGTGATTG TAGCCCTACC AGCATGTTAT AGGGCAAGAC 2502 CTTTGTGCTT TTGATCATTC TGGCCCATGA AAGCAACTTT GGTCTCCTTT CCCCTCCTGT 2562 CTTCCCGGTA TCCCTTGGAG TCTCACAAGG TTTACTTTGG TATGGTTCTC AGCACAAACC 2622 (2) INFORMATION FOR SEQ ID NO:13: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 657 amino acids (B) TYPE: amino acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: -112 - Glu Val Asp Val Leu Asp Val Asn Val Arg Gly Pro Asp Gly Cys Thr 1 5 10 15 Pro Leu Met Leu Ala Ser Leu Arg Gly Gly Ser Ser Asp Leu Ser Asp 20 25 30 Glu Asp Glu Asp Ala Glu Asp Ser Ser Ala Asn He He Thr Asp Leu 35 40 45 Val Tyr Gin Gly Ala Ser Leu Gin Ala Gin Thr Asp Arg Thr Gly Glu 50 55 60 Met Ala Leu His Leu Ala Ala Arg Tyr Ser Arg Ala Asp Ala Ala Lys 65 70 75 80 Arg Leu Leu Asp Ala Gly Ala Asp Ala Asn Ala Gin Asp Asn Met Gly 85 90 95 Arg Cys Pro Leu His Ala Ala Val Ala Ala Asp Ala Gin Gly Val Phe 100 105 110 Gin He Leu He Arg Asn Arg Val Thr Asp Leu Asp Ala Arg Met Asn 115 120 125 Asp Gly Thr Thr Pro Leu He Leu Ala Ala Arg Leu Ala Val Glu Gly 130 135 140 Met Val Ala Glu Leu He Asn Cys Gin Ala Asp Val Asn Ala Val Asp 145 150 155 160 Asp His Gly Lys Ser Ala Leu His Trp Ala Ala Ala Val Asn Asn Val 165 170 175 Glu Ala Thr Leu Leu Leu Leu Lys Asn Gly Ala Asn Arg Asp Met Gin 180 185 190 Asp Asn Lys Glu Glu Thr Pro Leu Phe Leu Ala Ala Arg Glu Gly Ser 195 200 205 Tyr Glu Ala Ala Lys He Leu Leu Asp His Phe Ala Asn Arg Asp He 210 215 220 Thr Asp His Met Asp Arg Leu Pro Arg Asp Val Ala Arg Asp Arg Met 225 230 235 240 His His Asp He Val Arg Leu Leu Asp Glu Tyr Asn Val Thr Pro Ser 245 250 255 Pro Pro Gly Thr Val Leu Thr Ser Ala Leu Ser Pro Val He Cys Gly 260 265 270 Pro Asn Arg Ser Phe Leu Ser Leu Lys His Thr Pro Met Gly Lys Lys 275 280 285 Ser Arg Arg Pro Ser Ala Lys Ser Thr Met Pro Thr Ser Leu Pro Asn 290 295 300 Leu Ala Lys Glu Ala Lys Asp Ala Lys Gly Ser Arg Arg Lys Lys Ser 305 310 315 320 Leu Ser Glu Lys Val Gin Leu Ser Glu Ser Ser Val Thr Leu Ser Pro 325 330 335 Val Asp Ser Leu Glu Ser Pro His Thr Tyr Val Ser Asp Thr Thr Ser 340 345 350 Ser Pro Met He Thr Ser Pro Gly He Leu Gin Ala Ser Pro Asn Pro -113- 355 360 365 Met Leu Ala Thr Ala Ala Pro Pro Ala Pro Val His Ala Gin His Ala 370 375 380 Leu Ser Phe Ser Asn Leu His Glu Met Gin Pro Leu Ala His Gly Ala 385 390 395 400 Ser Thr Val Leu Pro Ser Val Ser Gin Leu Leu Ser His His His He 405 410 415 Val Ser Pro Gly Ser Gly Ser Ala Gly Ser Leu Ser Arg Leu His Pro 420 425 430 Val Pro Val Pro Ala Asp Trp Met Asn Arg Met Glu Val Asn Glu Thr 435 440 445 Gin Tyr Asn Glu Met Phe Gly Met Val Leu Ala Pro Ala Glu Gly Thr 450 455 460 His Pro Gly He Ala Pro Gin Ser Arg Pro Pro Glu Gly Lys His He 465 470 475 480 Thr Thr Pro Arg Glu Pro Leu Pro Pro He Val Thr Phe Gin Leu He 485 490 495 Pro Lys Gly Ser He Ala Gin Pro Ala Gly Ala Pro Gin Pro Gin Ser 500 505 510 Thr Cys Pro Pro Ala Val Ala Gly Pro Leu Pro Thr Met Tyr Gin He 515 520 525 Pro Glu Met Ala Arg Leu Pro Ser Val Ala Phe Pro Thr Ala Met Met 530 535 540 Pro Gin Gin Asp Gly Gin Val Ala Gin Thr He Leu Pro Ala Tyr His 545 550 555 560 Pro Phe Pro Ala Ser Val Gly Lys Tyr Pro Thr Pro Pro Ser Gin His 565 570 575 Ser Tyr Ala Ser Ser Asn Ala Ala Glu Arg Thr Pro Ser His Ser Gly 580 585 590 His Leu Gin Gly Glu His Pro Tyr Leu Thr Pro Ser Pro Glu Ser Pro 595 600 605 Asp Gin Trp Ser Ser Ser Ser Pro His Ser Ala Ser Asp Trp Ser Asp 610 615 620 Val Thr Thr Ser Pro Thr Pro Gly Gly Ala Gly Gly Gly Gin Arg Gly 625 630 635 640 Pro Gly Thr His Met Ser Glu Pro Pro His Asn Asn Met Gin Val Tyr 645 650 655 Ala (2) INFORMATION FOR SEQ ID NO:14: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 77 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: unknown -114- (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: Glu Asp He Asp Glu Cys Asp Gin Gly Ser Pro Cys Glu His Asn Gly 1 5 10 15 He Cys Val Asn Thr Pro Gly Ser Tyr Arg Cys Asn Cys Ser Gin Gly 20 25 30 Phe Thr Gly Pro Arg Cys Glu Thr Asn He Asn Glu Cys Glu Ser His 35 40 45 Pro Cys Gin Asn Glu Gly Ser Cys Leu Asp Asp Pro Gly Thr Phe Arg 50 55 60 Cys Val Cys Met Pro Gly Phe Thr Gly Thr Gin Cys Glu 65 70 75 (2) INFORMATION FOR SEQ ID NO:15: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 78 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15: Asn Asp Val Asp Glu Cys Ser Leu Gly Ala Asn Pro Cys Glu His Gly 1 5 10 15 Gly Arg Cys Thr Asn Thr Leu Gly Ser Phe Gin Cys Asn Cys Pro Gin 20 25 30 Gly Tyr Ala Gly Pro Arg Cys Glu He Asp Val Asn Glu Cys Leu Ser 35 40 45 Asn Pro Cys Gin Asn Asp Ser Thr Cys Leu Asp Gin He Gly Glu Phe 50 55 60 Gin Cys He Cys Met Pro Gly Tyr Glu Gly Leu Tyr Cys Glu 65 70 75 (2) INFORMATION FOR SEQ ID NO:16: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 654 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: Thr Pro Pro Gin Gly Glu He Glu Ala Asp Cys Met Asp Val Asn Val -115- 1 5 10 15 Arg Gly Pro Asp Gly Phe Thr Pro Leu Met He Ala Ser Cys Ser Gly 20 25 30 Gly Gly Leu Glu Thr Gly Asn Ser Glu Glu Glu Glu Asp Ala Ser Ala 35 40 45 Asn Met He Ser Asp Phe He Gly Gin Gly Ala Gin Leu His Asn Gin 50 55 60 Thr Asp Arg Thr Gly Glu Thr Ala Leu His Leu Ala Ala Arg Tyr Ala 65 70 75 80 Arg Ala Asp Ala Ala Lys Arg Leu Leu Glu Ser Ser Ala Asp Ala Asn 85 90 95 Val Gin Asp Asn Met Gly Arg Thr Pro Leu His Ala Ala Val Ala Ala 100 105 110 Asp Ala Gin Gly Val Phe Gin He Leu He Arg Asn Arg Ala Thr Asp 115 120 125 Leu Asp Ala Arg Met Phe Asp Gly Thr Thr Pro Leu He Leu Ala Ala 130 135 140 Arg Leu Ala Val Glu Gly Met Val Glu Glu Leu He Asn Ala His Ala 145 150 155 160 Asp Val Asn Ala Val Asp Glu Phe Gly Lys Ser Ala Leu His Trp Ala 165 170 175 Ala Ala Val Asn Asn Val Asp Ala Ala Ala Val Leu Leu Lys Asn Ser 180 185 190 Ala Asn Lys Asp Met Gin Asn Asn Lys Glu Glu Thr Ser Leu Phe Leu 195 200 205 Ala Ala Arg Glu Gly Ser Tyr Glu Thr Ala Lys Val Leu Leu Asp His 210 215 220 Tyr Ala Asn Arg Asp He Thr Asp His Met Asp Arg Leu Pro Arg Asp 225 230 235 240 He Ala Gin Glu Arg Met His His Asp He Val His Leu Leu Asp Glu 245 250 255 Tyr Asn Leu Val Lys Ser Pro Thr Leu His Asn Gly Pro Leu Gly Ala 260 265 270 Thr Thr Leu Ser Pro Pro He Cys Ser Pro Asn Gly Tyr Met Gly Asn 275 280 285 Met Lys Pro Ser Val Gin Ser Lys Lys Ala Arg Lys Pro Ser He Lys 290 295 300 Gly Asn Gly Cys Lys Glu Ala Lys Glu Leu Lys Ala Arg Arg Lys Lys 305 310 315 320 Ser Gin Asp Gly Lys Thr Thr Leu Leu Asp Ser Gly Ser Ser Gly Val 325 330 335 Leu Ser Pro Val Asp Ser Leu Glu Ser Thr His Gly Tyr Leu Ser Asp 340 345 350 Val Ser Ser Pro Pro Leu Met Thr Ser Pro Phe Gin Gin Ser Pro Ser 355 360 365 -116- Met Pro Leu Asn His Leu Thr Ser Met Pro Glu Ser Gin Leu Gly Met 370 375 380 Asn His He Asn Met Ala Thr Lys Gin Glu Met Ala Ala Gly Ser Asn 385 390 395 400 Arg Met Ala Phe Asp Ala Met Val Pro Arg Leu Thr His Leu Asn Ala 405 410 415 Ser Ser Pro Asn Thr He Met Ser Asn Gly Ser Met His Phe Thr Val 420 425 430 Gly Gly Ala Pro Thr Met Asn Ser Gin Cys Asp Trp Leu Ala Arg Leu 435 440 445 Gin Asn Gly Met Val Gin Asn Gin Tyr Asp Pro He Arg Asn Gly He 450 455 460 Gin Gin Gly Asn Ala Gin Gin Ala Gin Ala Leu Gin His Gly Leu Met 465 470 475 480 Thr Ser Leu His Asn Gly Leu Pro Ala Thr Thr Leu Ser Gin Met Met 485 490 495 Thr Tyr Gin Ala Met Pro Asn Thr Arg Leu Ala Asn Gin Pro His Leu 500 505 510 Met Gin Ala Gin Gin Met Gin Gin Gin Gin Asn Leu Gin Leu His Gin 515 520 525 Ser Met Gin Gin Gin His His Asn Ser Ser Thr Thr Ser Thr His He 530 535 540 Asn Ser Pro Phe Cys Ser Ser Asp He Ser Gin Thr Asp Leu Gin Gin 545 550 555 560 Met Ser Ser Asn Asn He His Ser Val Met Pro Gin Asp Thr Gin He 565 570 575 Phe Ala Ala Ser Leu Pro Ser Asn Leu Thr Gin Ser Met Thr Thr Ala 580 585 590 Gin Phe Leu Thr Pro Pro Ser Gin His Ser Tyr Ser Ser Pro Met Asp 595 600 605 Asn Thr Pro Ser His Gin Leu Gin Val Pro Asp His Pro Phe Leu Thr 610 615 620 Pro Ser Pro Glu Ser Pro Asp Gin Trp Ser Ser Ser Ser Pro His Ser 625 630 635 640 Asn Met Ser Asp Trp Ser Glu Gly He Ser Ser Pro Pro Thr 645 650 (2) INFORMATION FOR SEQ ID NO:17: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 666 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 17: -117- Thr Pro Pro Gin Gly Glu Val Asp Ala Asp Cys Met Asp Val Asn Val 1 5 10 15 Arg Gly Pro Asp Gly Phe Thr Pro Leu Met He Ala Ser Cys Ser Gly 20 25 30 Gly Gly Leu Glu Thr Gly Asn Ser Glu Glu Glu Glu Asp Ala Pro Ala 35 40 45 Val He Ser Asp Phe He Tyr Gin Gly Ala Ser Leu His Asn Gin Thr 50 55 60 Asp Arg Thr Gly Glu Thr Ala Leu His Leu Ala Ala Arg Tyr Ser Arg 65 70 75 80 Ser Asp Ala Ala Lys Arg Leu Leu Glu Ala Ser Ala Asp Ala Asn He 85 90 95 Gin Asp Asn Met Gly Arg Thr Pro Leu His Ala Ala Val Ser Ala Asp 100 105 110 Ala Gin Gly Val Phe Gin He Leu Leu Arg Asn Arg Ala Thr Asp Leu 115 120 125 Asp Ala Arg Met His Asp Gly Thr Thr Pro Leu He Leu Ala Ala Arg 130 135 140 Leu Ala Val Glu Gly Met Leu Glu Asp Leu He Asn Ser His Ala Asp 145 150 155 160 Val Asn Ala Val Asp Asp Leu Gly Lys Ser Ala Leu His Trp Ala Ala 165 170 175 Ala Val Asn Asn Val Asp Ala Ala Val Val Leu Leu Lys Asn Gly Ala 180 185 190 Asn Lys Asp Met Gin Asn Asn Lys Glu Glu Thr Pro Leu Phe Leu Ala 195 200 205 Ala Arg Glu Gly Ser Tyr Glu Thr Ala Lys Val Leu Leu Asp His Phe 210 215 220 Ala Asn Arg Asp He Thr Asp His Met Asp Arg Leu Pro Arg Asp He 225 230 235 240 Ala Gin Glu Arg Met His His Asp He Val Arg Leu Leu Asp Glu Tyr 245 250 255 Asn Leu Val Arg Ser Pro Gin Leu His Gly Thr Ala Leu Gly Gly Thr 260 265 270 Pro Thr Leu Ser Pro Thr Leu Cys Ser Pro Asn Gly Tyr Leu Gly Asn 275 280 285 Leu Lys Ser Ala Thr Gin Gly Lys Lys Ala Arg Lys Pro Ser Thr Lys 290 295 300 Gly Leu Ala Cys Ser Ser Lys Glu Ala Lys Asp Leu Lys Ala Arg Arg 305 310 315 320 Lys Lys Ser Gin Asp Gly Lys Gly Cys Leu Leu Asp Ser Ser Ser Met 325 330 335 Leu Ser Pro Val Asp Ser Leu Glu Ser Pro His Gly Tyr Leu Ser Asp 340 345 350 Val Ala Ser Pro Pro Leu Pro Ser Pro Phe Gin Gin Ser Pro Ser Met -118- 355 360 365 Pro Leu Ser His Leu Pro Gly Met Pro Asp Thr His Leu Gly He Ser 370 375 380 His Leu Asn Val Ala Ala Lys Pro Glu Met Ala Ala Leu Ala Gly Gly 385 390 395 400 Ser Arg Leu Ala Phe Glu Pro Pro Pro Pro Arg Leu Ser His Leu Pro 405 410 415 Val Ala Ser Ser Ala Ser Thr Val Leu Ser Thr Asn Gly Thr Gly Ala 420 425 430 Met Asn Phe Thr Val Gly Ala Pro Ala Ser Leu Asn Gly Gin Cys Glu 435 440 445 Trp Leu Pro Arg Leu Gin Asn Gly Met Val Pro Ser Gin Tyr Asn Pro 450 455 460 Leu Arg Pro Gly Val Thr Pro Gly Thr Leu Ser Thr Gin Ala Ala Gly 465 470 475 480 Leu Gin His Gly Met Met Ser Pro He His Ser Ser Leu Ser Thr Asn 485 490 495 Thr Leu Ser Pro He He Tyr Gin Gly Leu Pro Asn Thr Arg Leu Ala 500 505 510 Thr Gin Pro His Leu Val Gin Thr Gin Gin Val Gin Pro Gin Asn Leu 515 520 525 Gin He Gin Pro Gin Asn Leu Gin Pro Pro Ser Gin Pro His Leu Ser 530 535 540 Val Ser Ser Ala Ala Asn Gly His Leu Gly Arg Ser Phe Leu Ser Gly 545 550 555 560 Glu Pro Ser Gin Ala Asp Val Gin Pro Leu Gly Pro Ser Ser Leu Pro 565 570 575 Val His Thr He Leu Pro Gin Glu Ser Gin Ala Leu Pro Thr Ser Leu 580 585 590 Pro Ser Ser Met Val Pro Pro Met Thr Thr Thr Gin Phe Leu Thr Pro 595 600 605 Pro Ser Gin His Ser Tyr Ser Ser Ser Pro Val Asp Asn Thr Pro Ser 610 615 620 His Gin Leu Gin Val Pro Glu His Pro Phe Leu Thr Pro Ser Pro Glu 625 630 635 640 Ser Pro Asp Gin Trp Ser Ser Ser Ser Arg His Ser Asn He Ser Asp 645 650 655 Trp Ser Glu Gly He Ser Ser Pro Pro Thr 660 665 (2) INFORMATION FOR SEQ ID NO:18: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 681 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: unknown -119- (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18: Thr Pro Pro Gin Gly Glu Val Asp Ala Asp Cys Met Asp Val Asn Val 1 5 10 15 Arg Gly Pro Asp Gly Phe Thr Pro Leu Met He Ala Ser Cys Ser Gly 20 25 30 Gly Gly Leu Glu Thr Gly Asn Ser Glu Glu Glu Glu Asp Ala Pro Ala 35 40 45 Val He Ser Asp Phe He Tyr Gin Gly Ala Ser Leu His Asn Gin Thr 50 55 60 Asp Arg Thr Gly Glu Thr Ala Leu His Leu Ala Ala Arg Tyr Ser Arg 65 70 75 80 Ser Asp Ala Ala Lys Arg Leu Leu Glu Ala Ser Ala Asp Ala Asn He 85 90 95 Gin Asp Asn Met Gly Arg Thr Pro Leu His Ala Ala Val Ser Ala Asp 100 105 110 Ala Gin Gly Val Phe Gin He Leu He Arg Asn Arg Ala Thr Asp Leu 115 120 125 Asp Ala Arg Met His Asp Gly Thr Thr Pro Leu He Leu Ala Ala Arg 130 135 140 Leu Ala Val Glu Gly Met Leu Glu Asp Leu He Asn Ser His Ala Asp 145 150 155 160 Val Asn Ala Val Asp Asp Leu Gly Lys Ser Ala Leu His Trp Ala Ala 165 170 175 Ala Val Asn Asn Val Asp Ala Ala Val Val Leu Leu Lys Asn Gly Ala 180 185 190 Asn Lys Asp Met Gin Asn Asn Arg Glu Glu Thr Pro Leu Phe Leu Ala 195 200 205 Ala Arg Glu Gly Ser Tyr Glu Thr Ala Lys Val Leu Leu Asp His Phe 210 215 220 Ala Asn Arg Asp He Thr Asp His Met Asp Arg Leu Pro Arg Asp He 225 230 235 240 Ala Gin Glu Arg Met His His Asp He Val Arg Leu Leu Asp Glu Tyr 245 250 255 Asn Leu Val Arg Ser Pro Gin Leu His Gly Ala Pro Leu Gly Gly Thr 260 265 270 Pro Thr Leu Ser Pro Pro Leu Cys Ser Pro Asn Gly Tyr Leu Gly Ser 275 280 285 Leu Lys Pro Gly Val Gin Gly Lys Lys Val Arg Lys Pro Ser Ser Lys 290 295 300 Gly Leu Ala Cys Gly Ser Lys Glu Ala Lys Asp Leu Lys Ala Arg Arg 305 310 315 320 Lys Lys Ser Gin Asp Gly Lys Gly Cys Leu Leu Asp Ser Ser Gly Met -120- 325 330 335 Leu Ser Pro Val Asp Ser Leu Glu Ser Pro His Gly Tyr Leu Ser Asp 340 345 350 Val Ala Ser Pro Pro Leu Leu Pro Ser Pro Phe Gin Gin Ser Pro Ser 355 360 365 Val Pro Leu Asn His Leu Pro Gly Met Pro Asp Thr His Leu Gly He 370 375 380 Gly His Leu Asn Val Ala Ala Lys Pro Glu Met Ala Ala Leu Gly Gly 385 390 395 400 Gly Gly Arg Leu Ala Phe Glu Thr Gly Pro Pro Arg Leu Ser His Leu 405 410 415 Pro Val Ala Ser Gly Thr Ser Thr Val Leu Gly Ser Ser Ser Gly Gly 420 425 430 Ala Leu Asn Phe Thr Val Gly Gly Ser Thr Ser Leu Asn Gly Gin Cys 435 440 445 Glu Trp Leu Ser Arg Leu Gin Ser Gly Met Val Pro Asn Gin Tyr Asn 450 455 460 Pro Leu Arg Gly Ser Val Ala Pro Gly Pro Leu Ser Thr Gin Ala Pro 465 470 475 480 Ser Leu Gin His Gly Met Val Gly Pro Leu His Ser Ser Leu Ala Ala 485 490 495 Ser Ala Leu Ser Gin Met Met Ser Tyr Gin Gly Leu Pro Ser Thr Arg 500 505 510 Leu Ala Thr Gin Pro His Leu Val Gin Thr Gin Gin Val Gin Pro Gin 515 520 525 v Asn Leu Gin Met Gin Gin Gin Asn Leu Gin Pro Ala Asn He Gin Gin 530 535 540 Gin Gin Ser Leu Gin Pro Pro Pro Pro Pro Pro Gin Pro His Leu Gly 545 550 555 560 Val Ser Ser Ala Ala Ser Gly His Leu Gly Arg Ser Phe Leu Ser Gly 565 570 575 Glu Pro Ser Gin Ala Asp Val Gin Pro Leu Gly Pro Ser Ser Leu Ala 580 585 590 Val His Thr He Leu Pro Gin Glu Ser Pro Ala Leu Pro Thr Ser Leu 595 600 605 Pro Ser Ser Leu Val Pro Pro Val Thr Ala Ala Gin Phe Leu Thr Pro 610 615 620 Pro Ser Gin His Ser Tyr Ser Ser Pro Val Glu Asn Thr Pro Ser His 625 630 635 640 Gin Leu Gin Val Pro Glu His Pro Phe Leu Thr Pro Ser Pro Glu Ser 645 650 655 Pro Asp Gin Trp Ser Ser Ser Ser Pro His Ser Asn Val Ser Asp Trp 660 665 670 Ser Glu Gly Val Ser Ser Pro Pro Thr 675 680 -121- (2) INFORMATION FOR SEQ ID NO:19: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2471 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: Met Pro Ala Leu Arg Pro Ala Leu Leu Trp Ala Leu Leu Ala Leu Trp 1 5 10 15 Leu Cys Cys Ala Ala Pro Ala His Ala Leu Gin Cys Arg Asp Gly Tyr 20 25 30 Glu Pro Cys Val Asn Glu Gly Met Cys Val Thr Tyr His Asn Gly Thr 35 40 45 Gly Tyr Cys Lys Cys Pro Glu Gly Phe Leu Gly Glu Tyr Cys Gin His 50 55 60 Arg Asp Pro Cys Glu Lys Asn Arg Cys Gin Asn Gly Gly Thr Cys Val 65 70 75 80 Ala Gin Ala Met Leu Gly Lys Ala Thr Cys Arg Cys Ala Ser Gly Phe 85 90 95 Thr Gly Glu Asp Cys Gin Tyr Ser Thr Ser His Pro Cys Phe Val Ser 100 105 110 Arg Pro Cys Leu Asn Gly Gly Thr Cys His Met Leu Ser Arg Asp Thr 115 120 125 Tyr Glu Cys Thr Cys Gin Val Gly Phe Thr Gly Lys Glu Cys Gin Trp 130 135 140 Thr Asp Ala Cys Leu Ser His Pro Cys Ala Asn Gly Ser Thr Cys Thr 145 150 155 160 Thr Val Ala Asn Gin Phe Ser Cys Lys Cys Leu Thr Gly Phe Thr Gly 165 170 175 Gin Lys Cys Glu Thr Asp Val Asn Glu Cys Asp He Pro Gly His Cys 180 185 190 Gin His Gly Gly Thr Cys Leu Asn Leu Pro Gly Ser Tyr Gin Cys Gin 195 200 205 Cys Pro Gin Gly Phe Thr Gly Gin Tyr Cys Asp Ser Leu Tyr Val Pro 210 215 220 Cys Ala Pro Ser Pro Cys Val Asn Gly Gly Thr Cys Arg Gin Thr Gly 225 230 235 240 Asp Phe Thr Phe Glu Cys Asn Cys Leu Pro Gly Phe Glu Gly Ser Thr 245 250 255 Cys Glu Arg Asn He Asp Asp Cys Pro Asn His Arg Cys Gin Asn Gly 260 265 270 Gly Val Cys Val Asp Gly Val Asn Thr Tyr Asn Cys Arg Cys Pro Pro 275 280 285 -122- Gln Trp Thr Gly Gin Phe Cys Thr Glu Asp Val Asp Glu Cys Leu Leu 290 295 300 Gin Pro Asn Ala Cys Gin Asn Gly Gly Thr Cys Ala Asn Arg Asn Gly 305 310 315 320 Gly Tyr Gly Cys Val Cys Val Asn Gly Trp Ser Gly Asp Asp Cys Ser 325 330 335 Glu Asn He Asp Asp Cys Ala Phe Ala Ser Cys Thr Pro Gly Ser Thr 340 345 350 Cys He Asp Arg Val Ala Ser Phe Ser Cys Met Cys Pro Glu Gly Lys 355 360 365 Ala Gly Leu Leu Cys His Leu Asp Asp Ala Cys He Ser Asn Pro Cys 370 375 380 His Lys Gly Ala Leu Cys Asp Thr Asn Pro Leu Asn Gly Gin Tyr He 385 390 395 400 Cys Thr Cys Pro Gin Gly Tyr Lys Gly Ala Asp Cys Thr Glu Asp Val 405 410 415 Asp Glu Cys Ala Met Ala Asn Ser Asn Pro Cys Glu His Ala Gly Lys 420 425 430 Cys Val Asn Thr Asp Gly Ala Phe His Cys Glu Cys Leu Lys Gly Tyr 435 440 445 Ala Gly Pro Arg Cys Glu Met Asp He Asn Glu Cys His Ser Asp Pro 450 455 460 Cys Gin Asn Asp Ala Thr Cys Leu Asp Lys He Gly Gly Phe Thr Cys 465 470 475 480 Leu Cys Met Pro Gly Phe Lys Gly Val His Cys Glu Leu Glu He Asn 485 490 495 Glu Cys Gin Ser Asn Pro Cys Val Asn Asn Gly Gin Cys Val Asp Lys 500 505 510 Val Asn Arg Phe Gin Cys Leu Cys Pro Pro Gly Phe Thr Gly Pro Val 515 520 525 Cys Gin He Asp He Asp Asp Cys Ser Ser Thr Pro Cys Leu Asn Gly 530 535 540 Ala Lys Cys He Asp His Pro Asn Gly Tyr Glu Cys Gin Cys Ala Thr 545 550 555 560 Gly Phe Thr Gly Val Leu Cys Glu Glu Asn He Asp Asn Cys Asp Pro 565 570 575 Asp Pro Cys His His Gly Gin Cys Gin Asp Gly He Asp Ser Tyr Thr 580 585 590 Cys He Cys Asn Pro Gly Tyr Met Gly Ala He Cys Ser Asp Gin He 595 600 605 Asp Glu Cys Tyr Ser Ser Pro Cys Leu Asn Asp Gly Arg Cys He Asp 610 615 620 Leu Val Asn Gly Tyr Gin Cys Asn Cys Gin Pro Gly Thr Ser Gly Val 625 630 635 640 Asn Cys Glu He Asn Phe Asp Asp Cys Ala Ser Asn Pro Cys He His -123- 645 650 655 Gly He Cys Met Asp Gly He Asn Arg Tyr Ser Cys Val Cys Ser Pro 660 665 670 Gly Phe Thr Gly Gin Arg Cys Asn He Asp He Asp Glu Cys Ala Ser 675 680 685 Asn Pro Cys Arg Lys Gly Ala Thr Cys He Asn Gly Val Asn Gly Phe 690 695 700 Arg Cys He Cys Pro Glu Gly Pro His His Pro Ser Cys Tyr Ser Gin 705 710 715 720 Val Asn Glu Cys Leu Ser Asn Pro Cys He His Gly Asn Cys Thr Gly 725 730 735 Gly Leu Ser Gly Tyr Lys Cys Leu Cys Asp Ala Gly Trp Val Gly He 740 745 750 Asn Cys Glu Val Asp Lys Asn Glu Cys Leu Ser Asn Pro Cys Gin Asn 755 760 765 Gly Gly Thr Cys Asp Asn Leu Val Asn Gly Tyr Arg Cys Thr Cys Lys 770 775 780 Lys Gly Phe Lys Gly Tyr Asn Cys Gin Val Asn He Asp Glu Cys Ala 785 790 795 800 Ser Asn Pro Cys Leu Asn Gin Gly Thr Cys Phe Asp Asp He Ser Gly 805 810 815 Tyr Thr Cys His Cys Val Leu Pro Tyr Thr Gly Lys Asn Cys Gin Thr 820 825 830 Val Leu Ala Pro Cys Ser Pro Asn Pro Cys Glu Asn Ala Ala Val Cys 835 840 845 Lys Glu Ser Pro Asn Phe Glu Ser Tyr Thr Cys Leu Cys Ala Pro Gly 850 855 860 Trp Gin Gly Gin Arg Cys Thr He Asp He Asp Glu Cys He Ser Lys 865 870 875 880 Pro Cys Met Asn His Gly Leu Cys His Asn Thr Gin Gly Ser Tyr Met 885 890 895 Cys Glu Cys Pro Pro Gly Phe Ser Gly Met Asp Cys Glu Glu Asp He 900 905 910 Asp Asp Cys Leu Ala Asn Pro Cys Gin Asn Gly Gly Ser Cys Met Asp 915 920 925 Gly Val Asn Thr Phe Ser Cys Leu Cys Leu Pro Gly Phe Thr Gly Asp 930 935 940 Lys Cys Gin Thr Asp Met Asn Glu Cys Leu Ser Glu Pro Cys Lys Asn 945 950 955 960 Gly Gly Thr Cys Ser Asp Tyr Val Asn Ser Tyr Thr Cys Lys Cys Gin 965 970 975 Ala Gly Phe Asp Gly Val His Cys Glu Asn Asn He Asn Glu Cys Thr 980 985 990 Glu Ser Ser Cys Phe Asn Gly Gly Thr Cys Val Asp Gly He Asn Ser 995 1000 1005 -124- Phe Ser Cys Leu Cys Pro Val Gly Phe Thr Gly Ser Phe Cys Leu His 1010 1015 1020 Glu He Asn Glu Cys Ser Ser His Pro Cys Leu Asn Glu Gly Thr Cys 1025 1030 1035 1040 Val Asp Gly Leu Gly Thr Tyr Arg Cys Ser Cys Pro Leu Gly Tyr Thr 1045 1050 1055 Gly Lys Asn Cys Gin Thr Leu Val Asn Leu Cys Ser Arg Ser Pro Cys 1060 1065 1070 Lys Asn Lys Gly Thr Cys Val Gin Lys Lys Ala Glu Ser Gin Cys Leu 1075 1080 1085 Cys Pro Ser Gly Trp Ala Gly Ala Tyr Cys Asp Val Pro Asn Val Ser 1090 1095 1100 Cys Asp He Ala Ala Ser Arg Arg Gly Val Leu Val Glu His Leu Cys 1105 1110 1115 1120 Gin His Ser Gly Val Cys He Asn Ala Gly Asn Thr His Tyr Cys Gin 1125 1130 1135 Cys Pro Leu Gly Tyr Thr Gly Ser Tyr Cys Glu Glu Gin Leu Asp Glu 1140 1145 1150 Cys Ala Ser Asn Pro Cys Gin Hi Gly Ala Thr Cys Ser Asp Phe He 1155 1160 1165 Gly Gly Tyr Arg Cys Glu Cys Val Pro Gly Tyr Gin Gly Val Asn Cys 1170 1175 1180 Glu Tyr Glu Val Asp Glu Cys Gin Asn Gin Pro Cys Gin Asn Gly Gly 1185 1190 1195 1200 Thr Cys He Asp Leu Val Asn His Phe Lys Cys Ser Cys Pro Pro Gly 1205 1210 1215 Thr Arg Gly Leu Leu Cys Glu Glu Asn He Asp Asp Cys Ala Arg Gly 1220 1225 1230 Pro His Cys Leu Asn Gly Gly Gin Cys Met Asp Arg He Gly Gly Tyr 1235 1240 1245 Ser Cys Arg Cys Leu Pro Gly Phe Ala Gly Glu Arg Cys Glu Gly Asp 1250 1255 1260 He Asn Glu Cys Leu Ser Asn Pro Cys Ser Ser Glu Gly Ser Leu Asp 1265 1270 1275 1280 Cys He Gin Leu Thr Asn Asp Tyr Leu Cys Val Cys Arg Ser Ala Phe 1285 1290 1295 Thr Gly Arg His Cys Glu Thr Phe Val Asp Val Cys Pro Gin Met Pro 1300 1305 1310 Cys Leu Asn Gly Gly Thr Cys Ala Val Ala Ser Asn Met Pro Asp Gly 1315 1320 1325 Phe He Cys Arg Cys Pro Pro Gly Phe Ser Gly Ala Arg Cys Gin Ser 1330 1335 1340 Ser Cys Gly Gin Val Lys Cys Arg Lys Gly Glu Gin Cys Val His Thr 1345 1350 1355 1360 Ala Ser Gly Pro Arg Cys Phe Cys Pro Ser Pro Arg Asp Cys Glu Ser -125- 1365 1370 1375 Gly Cys Ala Ser Ser Pro Cys Gin His Gly Gly Ser Cys His Pro Gin 1380 1385 1390 Arg Gin Pro Pro Tyr Tyr Ser Cys Gin Cys Ala Pro Pro Phe Ser Gly 1395 1400 1405 Ser Arg Cys Glu Leu Tyr Thr Ala Pro Pro Ser Thr Pro Pro Ala Thr 1410 1415 1420 Cys Leu Ser Gin Tyr Cys Ala Asp Lys Ala Arg Asp Gly Val Cys Asp 1425 1430 1435 1440 Glu Ala Cys Asn Ser His Ala Cys Gin Trp Asp Gly Gly Asp Cys Ser 1445 1450 1455 Leu Thr Met Glu Asn Pro Trp Ala Asn Cys Ser Ser Pro Leu Pro Cys 1460 1465 1470 Trp Asp Tyr He Asn Asn Gin Cys Asp Glu Leu Cys Asn Thr Val Glu 1475 1480 1485 Cys Leu Phe Asp Asn Phe Glu Cys Gin Gly Asn Ser Lys Thr Cys Lys 1490 1495 1500 Tyr Asp Lys Tyr Cys Ala Asp His Phe Lys Asp Asn His Cys Asn Gin 1505 1510 1515 1520 Gly Cys Asn Ser Glu Glu Cys Gly Trp Asp Gly Leu Asp Cys Ala Ala 1525 1530 1535 Asp Gin Pro Glu Asn Leu Ala Glu Gly Thr Leu Val He Val Val Leu 1540 1545 1550 Met Pro Pro Glu Gin Leu Leu Gin Asp Ala Arg Ser Phe Leu Arg Ala 1555 1560 1565 Leu Gly Thr Leu Leu His Thr Asn Leu Arg He Lys Arg Asp Ser Gin 1570 1575 1580 Gly Glu Leu Met Val Tyr Pro Tyr Tyr Gly Glu Lys Ser Ala Ala Met 1585 1590 1595 1600 Lys Lys Gin Arg Met Thr Arg Arg Ser Leu Pro Gly Glu Gin Glu Gin 1605 1610 1615 Glu Val Ala Gly Ser Lys Val Phe Leu Glu He Asp Asn Arg Gin Cys 1620 1625 1630 Val Gin Asp Ser Asp His Cys Phe Lys Asn Thr Asp Ala Ala Ala Ala 1635 1640 1645 Leu Leu Ala Ser His Ala He Gin Gly Thr Leu Ser Tyr Pro Leu Val 1650 1655 1660 Ser Val Val Ser Glu Ser Leu Thr Pro Glu Arg Thr Gin Leu Leu Tyr 1665 1670 1675 1680 Leu Leu Ala Val Ala Val Val He He Leu Phe He He Leu Leu Gly 1685 1690 1695 Val He Met Ala Lys Arg Lys Arg Lys His Gly Ser Leu Trp Leu Pro 1700 1705 1710 Glu Gly Phe Thr Leu Arg Arg Asp Ala Ser Asn His Lys Arg Arg Glu 1715 1720 1725 -126- Pro Val Gly Gin Asp Ala Val Gly Leu Lys Asn Leu Ser Val Gin Val 1730 1735 1740 Ser Glu Ala Asn Leu He Gly Thr Gly Thr Ser Glu His Trp Val Asp 1745 1750 1755 1760 Asp Glu Gly Pro Gin Pro Lys Lys Val Lys Ala Glu Asp Glu Ala Leu 1765 1770 1775 Leu Ser Glu Glu Asp Asp Pro He Asp Arg Arg Pro Trp Thr Gin Gin 1780 1785 1790 His Leu Glu Ala Ala Asp He Arg Arg Thr Pro Ser Leu Ala Leu Thr 1795 1800 1805 Pro Pro Gin Ala Glu Gin Glu Val Asp Val Leu Asp Val Asn Val Arg 1810 1815 1820 Gly Pro Asp Gly Cys Thr Pro Leu Met Leu Ala Ser Leu Arg Gly Gly 1825 1830 1835 1840 Ser Ser Asp Leu Ser Asp Glu Asp Glu Asp Ala Glu Asp Ser Ser Ala 1845 1850 1855 Asn He He Thr Asp Leu Val Tyr Gin Gly Ala Ser Leu Gin Ala Gin 1860 1865 1870 Thr Asp Arg Thr Gly Glu Met Ala Leu His Leu Ala Ala Arg Tyr Ser 1875 1880 1885 Arg Ala Asp Ala Ala Lys Arg Leu Leu Asp Ala Gly Ala Asp Ala Asn 1890 1895 1900 Ala Gin Asp Asn Met Gly Arg Cys Pro Leu His Ala Ala Val Ala Ala 1905 1910 1915 1920 Asp Ala Gin Gly Val Phe Gin He Leu He Arg Asn Arg Val Thr Asp 1925 1930 1935 Leu Asp Ala Arg Met Asn Asp Gly Thr Thr Pro Leu He Leu Ala Ala 1940 1945 1950 Arg Leu Ala Val Glu Gly Met Val Ala Glu Leu He Asn Cys Gin Ala 1955 1960 1965 Asp Val Asn Ala Val Asp Asp His Gly Lys Ser Ala Leu His Trp Ala 1970 1975 1980 Ala Ala Val Asn Asn Val Glu Ala Thr Leu Leu Leu Leu Lys Asn Gly 1985 1990 1995 2000 Ala Asn Arg Asp Met Gin Asp Asn Lys Glu Glu Thr Pro Leu Phe Leu 2005 2010 2015 Ala Ala Arg Glu Gly Ser Tyr Glu Ala Ala Lys He Leu Leu Asp His 2020 2025 2030 Phe Ala Asn Arg Asp He Thr Asp His Met Asp Arg Leu Pro Arg Asp 2035 2040 2045 Val Ala Arg Asp Arg Met His His Asp He Val Arg Leu Leu Asp Glu 2050 2055 2060 Tyr Asn Val Thr Pro Ser Pro Pro Gly Thr Val Leu Thr Ser Ala Leu 2065 2070 2075 2080 Ser Pro Val He Cys Gly Pro Asn Arg Ser Phe Leu Ser Leu Lys His -127- 2085 2090 2095 Thr Pro Met Gly Lys Lys Ser Arg Arg Pro Ser Ala Lys Ser Thr Met 2100 2105 2110 Pro Thr Ser Leu Pro Asn Leu Ala Lys Glu Ala Lys Asp Ala Lys Gly 2115 2120 2125 Ser Arg Arg Lys Lys Ser Leu Ser Glu Lys Val Gin Leu Ser Glu Ser 2130 2135 2140 Ser Val Thr Leu Ser Pro Val Asp Ser Leu Glu Ser Pro His Thr Tyr 2145 2150 2155 2160 Val Ser Asp Thr Thr Ser Ser Pro Met He Thr Ser Pro Gly He Leu 2165 2170 2175 Gin Ala Ser Pro Asn Pro Met Leu Ala Thr Ala Ala Pro Pro Ala Pro 2180 ■ 2185 2190 Val His Ala Gin His Ala Leu Ser Phe Ser Asn Leu His Glu Met Gin 2195 2200 2205 Pro Leu Ala His Gly Ala Ser Thr Val Leu Pro Ser Val Ser Gin Leu 2210 2215 2220 Leu Ser His His His He Val Ser Pro Gly Ser Gly Ser Ala Gly Ser 2225 2230 2235 2240 Leu Ser Arg Leu His Pro Val Pro Val Pro Ala Asp Trp Met Asn Arg 2245 2250 2255 Met Glu Val Asn Glu Thr Gin Tyr Asn Glu Met Phe Gly Met Val Leu 2260 2265 2270 Ala Pro Ala Glu Gly Thr His Pro Gly He Ala Pro Gin Ser Arg Pro 2275 2280 2285 Pro Glu Gly Lys His He Thr Thr Pro Arg Glu Pro Leu Pro Pro He 2290 2295 2300 Val Thr Phe Gin Leu He Pro Lys Gly Ser He Ala Gin Pro Ala Gly 2305 2310 2315 2320 Ala Pro Gin Pro Gin Ser Thr Cys Pro Pro Ala Val Ala Gly Pro Leu 2325 2330 2335 Pro Thr Met Tyr Gin He Pro Glu Met Ala Arg Leu Pro Ser Val Ala 2340 2345 2350 Phe Pro Thr Ala Met Met Pro Gin Gin Asp Gly Gin Val Ala Gin Thr 2355 2360 2365 He Leu Pro Ala Tyr His Pro Phe Pro Ala Ser Val Gly Lys Tyr Pro 2370 2375 2380 Thr Pro Pro Ser Gin His Ser Tyr Ala Ser Ser Asn Ala Ala Glu Arg 2385 2390 2395 2400 Thr Pro Ser His Ser Gly His Leu Gin Gly Glu His Pro Tyr Leu Thr 2405 2410 2415 Pro Ser Pro Glu Ser Pro Asp Gin Trp Ser Ser Ser Ser Pro His Ser 2420 2425 2430 Ala Ser Asp Trp Ser Asp Val Thr Thr Ser Pro Thr Pro Gly Gly Ala 2435 2440 2445 -128- Gly Gly Gly Gin Arg Gly Pro Gly Thr His Met Ser Glu Pro Pro His 2450 2455 2460 Asn Asn Met Gin Val Tyr Ala 2465 2470 (2) INFORMATION FOR SEQ ID NO:20: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2556 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20: Met Pro Pro Leu Leu Ala Pro Leu Leu Cys Leu Ala Leu Leu Pro Ala 1 5 10 15 Leu Ala Ala Arg Gly Pro Arg Cys Ser Gin Pro Gly Glu Thr Cys Leu 20 25 30 Asn Gly Gly Lys Cys Glu Ala Ala Asn Gly Thr Glu Ala Cys Val Cys 35 40 45 Gly Gly Ala Phe Val Gly Pro Arg Cys Gin Asp Pro Asn Pro Cys Leu 50 55 60 Ser Thr Pro Cys Lys Asn Ala Gly Thr Cys His Val Val Asp Arg Arg 65 70 75 80 Gly Val Ala Asp Tyr Ala Cys Ser Cys Ala Leu Gly Phe Ser Gly Pro 85 90 95 Leu Cys Leu Thr Pro Leu Asp Asn Ala Cys Leu Thr Asn Pro Cys Arg 100 105 110 Asn Gly Gly Thr Cys Asp Leu Leu Thr Leu Thr Glu Tyr Lys Cys Arg 115 120 125 Cys Pro Pro Gly Trp Ser Gly Lys Ser Cys Gin Gin Ala Asp Pro Cys 130 135 140 Ala Ser Asn Pro Cys Ala Asn Gly Gly Gin Cys Leu Pro Phe Glu Ala 145 150 155 160 Ser Tyr He Cys His Cys Pro Pro Ser Phe His Gly Pro Thr Cys Arg 165 170 175 Gin Asp Val Asn Glu Cys Gly Gin Lys Pro Arg Leu Cys Arg His Gly 180 185 190 Gly Thr Cys His Asn Glu Val Gly Ser Tyr Arg Cys Val Cys Arg Ala 195 200 205 Thr His Thr Gly Pro Asn Cys Glu Arg Pro Tyr Val Pro Cys Ser Pro 210 215 220 Ser Pro Cys Gin Asn Gly Gly Thr Cys Arg Pro Thr Gly Asp Val Thr 225 230 235 240 His Glu Cys Ala Cys Leu Pro Gly Phe Thr Gly Gin Asn Cys Glu Glu 245 250 255 -129- Asn He Asp Asp Cys Pro Gly Asn Asn Cys Lys Asn Gly Gly Ala Cys 260 265 270 Val Asp Gly Val Asn Thr Tyr Asn Cys Pro Cys Pro Pro Glu Trp Thr 275 280 285 Gly Gin Tyr Cys .Thr Glu Asp Val Asp Glu Cys Gin Leu Met Pro Asn 290 295 300 Ala Cys Gin Asn Gly Gly Thr Cys His Asn Thr His Gly Gly Tyr Asn 305 310 315 320 Cys Val Cys Val Asn Gly Trp Thr Gly Glu Asp Cys Ser Glu Asn He 325 330 335 Asp Asp Cys Ala Ser Ala Ala Cys Phe His Gly Ala Thr Cys His Asp 340 345 350 Arg Val Ala Ser Phe Tyr Cys Glu Cys Pro His Gly Arg Thr Gly Leu 355 360 365 Leu Cys His Leu Asn Asp Ala Cys He Ser Asn Pro Cys Asn Glu Gly 370 375 380 Ser Asn Cys Asp Thr Asn Pro Val Asn Gly Lys Ala He Cys Thr Cys 385 390 395 400 Pro Ser Gly Tyr Thr Gly Pro Ala Cys Ser Gin Asp Val Asp Glu Cys 405 410 415 Ser Leu Gly Ala Asn Pro Cys Glu His Ala Gly Lys Cys He Asn Thr 420 425 430 Leu Gly Ser Phe Glu Cys Gin Cys Leu Gin Gly Tyr Thr Gly Pro Arg 435 440 445 Cys Glu He Asp Val Asn Glu Cys Val Ser Asn Pro Cys Gin Asn Asp 450 455 460 Ala Thr Cys Leu Asp Gin He Gly Glu Phe Gin Cys Met Cys Met Pro 465 470 475 480 Gly Tyr Glu Gly Val His Cys Glu Val Asn Thr Asp Glu Cys Ala Ser 485 490 495 Ser Pro Cys Leu His Asn Gly Arg Cys Leu Asp Lys He Asn Glu Phe 500 505 510 Gin Cys Glu Cys Pro Thr Gly Phe Thr Gly His Leu Cys Gin Tyr Asp 515 520 525 Val Asp Glu Cys Ala Ser Thr Pro Cys Lys Asn Gly Ala Lys Cys Leu 530 535 540 Asp Gly Pro Asn Thr Tyr Thr Cys Val Cys Thr Glu Gly Tyr Thr Gly 545 550 555 560 Thr His Cys Glu Val Asp He Asp Glu Cys Asp Pro Asp Pro Cys His 565 570 575 Tyr Gly Ser Cys Lys Asp Gly Val Ala Thr Phe Thr Cys Leu Cys Arg 580 585 590 Pro Gly Tyr Thr Gly His His Cys Glu Thr Asn He Asn Glu Cys Ser 595 600 605 Ser Gin Pro Cys Arg Leu Arg Gly Thr Cys Gin Asp Pro Asp Asn Ala -130- 610 615 620 Tyr Leu Cys Phe Cys Leu Lys Gly Thr Thr Gly Pro Asn Cys Glu He 625 630 635 640 Asn Leu Asp Asp Cys Ala Ser Ser Pro Cys Asp Ser Gly Thr Cys Leu 645 650 655 Asp Lys He Asp Gly Tyr Glu Cys Ala Cys Glu Pro Gly Tyr Thr Gly 660 665 670 Ser Met Cys Asn Ser Asn He Asp Glu Cys Ala Gly Asn Pro Cys His 675 680 685 Asn Gly Gly Thr Cys Glu Asp Gly He Asn Gly Phe Thr Cys Arg Cys 690 695 700 Pro Glu Gly Tyr His Asp Pro Thr Cys Leu Ser Glu Val Asn Glu Cys 705 710 715 720 Asn Ser Asn Pro Cys Val His Gly Ala Cys Arg Asp Ser Leu Asn Gly 725 730 735 Tyr Lys Cys Asp Cys Asp Pro Gly Trp Ser Gly Thr Asn Cys Asp He 740 745 750 Asn Asn Asn Glu Cys Glu Ser Asn Pro Cys Val Asn Gly Gly Thr Cys 755 760 765 Lys Asp Met Thr Ser Gly He Val Cys Thr Cys Arg Glu Gly Phe Ser 770 775 780 Gly Pro Asn Cys Gin Thr Asn He Asn Glu Cys Ala Ser Asn Pro Cys 785 790 795 800 Leu Asn Lys Gly Thr Cys He Asp Asp Val Ala Gly Tyr Lys Cys Asn 805 810 815 Cys Leu Leu Pro Tyr Thr Gly Ala Thr Cys Glu Val Val Leu Ala Pro 820 825 830 Cys Ala Pro Ser Pro Cys Arg Asn Gly Gly Glu Cys Arg Gin Ser Glu 835 840 845 Asp Tyr Glu Ser Phe Ser Cys Val Cys Pro Thr Ala Gly Ala Lys Gly 850 855 860 Gin Thr Cys Glu Val Asp He Asn Glu Cys Val Leu Ser Pro Cys Arg 865 870 875 880 His Gly Ala Ser Cys Gin Asn Thr His Gly Gly Tyr Arg Cys His Cys 885 890 895 Gin Ala Gly Tyr Ser Gly Arg Asn Cys Glu Thr Asp He Asp Asp Cys 900 905 910 Arg Pro Asn Pro Cys His Asn Gly Gly Ser Cys Thr Asp Gly He Asn 915 920 925 Thr Ala Phe Cys Asp Cys Leu Pro Gly Phe Arg Gly Thr Phe Cys Glu 930 935 940 Glu Asp He Asn Glu Cys Ala Ser Asp Pro Cys Arg Asn Gly Ala Asn 945 950 955 960 Cys Thr Asp Cys Val Asp Ser Tyr Thr Cys Thr Cys Pro Ala Gly Phe 965 970 975 -131- Ser Gly He His Cys Glu Asn Asn Thr Pro Asp Cys Thr Glu Ser Ser 980 985 990 Cys Phe Asn Gly Gly Thr Cys Val Asp Gly He Asn Ser Phe Thr Cys 995 1000 1005 Leu Cys Pro Pro Gly Phe Thr Gly Ser Tyr Cys Gin His Val Val Asn 1010 1015 1020 Glu Cys Asp Ser Arg Pro Cys Leu Leu Gly Gly Thr Cys Gin Asp Gly 1025 1030 1035 1040 Arg Gly Leu His Arg Cys Thr Cys Pro Gin Gly Tyr Thr Gly Pro Asn 1045 1050 1055 Cys Gin Asn Leu Val His Trp Cys Asp Ser Ser Pro Cys Lys Asn Gly 1060 1065 1070 Gly Lys Cys Trp Gin Thr His Thr Gin Tyr Arg Cys Glu Cys Pro Ser 1075 1080 1085 Gly Trp Thr Gly Leu Tyr Cys Asp Val Pro Ser Val Ser Cys Glu Val 1090 1095 1100 Ala Ala Gin Arg Gin Gly Val Asp Val Ala Arg Leu Cys Gin His Gly 1105 1110 1115 1120 Gly Leu Cys Val Asp Ala Gly Asn Thr His His Cys Arg Cys Gin Ala 1125 1130 1135 Gly Tyr Thr Gly Ser Tyr Cys Glu Asp Leu Val Asp Glu Cys Ser Pro 1140 1145 1150 Ser Pro Cys Gin Asn Gly Ala Thr Cys Thr Asp Tyr Leu Gly Gly Tyr 1155 1160 1165 Ser Cys Lys Cys Val Ala Gly Tyr His Gly Val Asn Cys Ser Glu Glu 1170 v 1175 1180 He Asp Glu Cys Leu Ser His Pro Cys Gin Asn Gly Gly Thr Cys Leu 1185 1190 1195 1200 Asp Leu Pro Asn Thr Tyr Lys Cys Ser Cys Pro Arg Gly Thr Gin Gly 1205 1210 1215 Val His Cys Glu He Asn Val Asp Asp Cys Asn Pro Pro Val Asp Pro 1220 1225 1230 Val Ser Arg Ser Pro Lys Cys Phe Asn Asn Gly Thr Cys Val Asp Gin 1235 1240 1245 Val Gly Gly Tyr Ser Cys Thr Cys Pro Pro Gly Phe Val Gly Glu Arg 1250 1255 1260 Cys Glu Gly Asp Val Asn Glu Cys Leu Ser Asn Pro Cys Asp Ala Arg 1265 1270 1275 1280 Gly Thr Gin Asn Cys Val Gin Arg Val Asn Asp Phe His Cys Glu Cys 1285 1290 1295 Arg Ala Gly His Thr Gly Arg Arg Cys Glu Ser Val He Asn Gly Cys 1300 1305 1310 Lys Gly Lys Pro Cys Lys Asn Gly Gly Thr Cys Ala Val Ala Ser Asn 1315 1320 1325 Thr Ala Arg Gly Phe He Cys Lys Cys Pro Ala Gly Phe Glu Gly Ala -132- 1330 1335 1340 Thr Cys Glu Asn Asp Ala Arg Thr Cys Gly Ser Leu Arg Cys Leu Asn 1345 1350 1355 1360 Gly Gly Thr Cys He Ser Gly Pro Arg Ser Pro Thr Cys Leu Cys Leu 1365 1370 1375 Gly Pro Phe Thr Gly Pro Glu Cys Gin Phe Pro Ala Ser Ser Pro Cys 1380 1385 1390 Leu Gly Gly Asn Pro Cys Tyr Asn Gin Gly Thr Cys Glu Pro Thr Ser 1395 1400 1405 Glu Ser Pro Phe Tyr Arg Cys Leu Cys Pro Ala Lys Phe Asn Gly Leu 1410 1415 1420 Leu Cys His He Leu Asp Tyr Ser Phe Gly Gly Gly Ala Gly Arg Asp 1425 1430 1435 1440 He Pro Pro Pro Leu He Glu Glu Ala Cys Glu Leu Pro Glu Cys Gin 1445 1450 1455 Glu Asp Ala Gly Asn Lys Val Cys Ser Leu Gin Cys Asn Asn His Ala 1460 1465 1470 Cys Gly Trp Asp Gly Gly Asp Cys Ser Leu Asn Phe Asn Asp Pro Trp 1475 1480 1485 Lys Asn Cys Thr Gin Ser Leu Gin Cys Trp Lys Tyr Phe Ser Asp Gly 1490 1495 1500 His Cys Asp Ser Gin Cys Asn Ser Ala Gly Cys Leu Phe Asp Gly Phe 1505 1510 1515 1520 Asp Cys Gin Arg Ala Glu Gly Gin Cys Asn Pro Leu Tyr Asp Gin Tyr 1525 1530 1535 Cys Lys Asp His Phe Ser Asp Gly His Cys Asp Gin Gly Cys Asn Ser 1540 1545 1550 Ala Glu Cys Glu Trp Asp Gly Leu Asp Cys Ala Glu His Val Pro Glu 1555 1560 1565 Arg Leu Ala Ala Gly Thr Leu Val Val Val Val Leu Met Pro Pro Glu 1570 1575 1580 Gin Leu Arg Asn Ser Ser Phe His Phe Leu Arg Glu Leu Ser Arg Val 1585 1590 1595 1600 Leu His Thr Asn Val Val Phe Lys Arg Asp Ala His Gly Gin Gin Met 1605 1610 1615 He Phe Pro Tyr Tyr Gly Arg Glu Glu Glu Leu Arg Lys His Pro He 1620 1625 1630 Lys Arg Ala Ala Glu Gly Trp Ala Ala Pro Asp Ala Leu Leu Gly Gin 1635 1640 1645 Val Lys Ala Ser Leu Leu Pro Gly Gly Ser Glu Gly Gly Arg Arg Arg 1650 1655 1660 Arg Glu Leu Asp Pro Met Asp Val Arg Gly Ser He Val Tyr Leu Glu 1665 1670 1675 1680 He Asp Asn Arg Gin Cys Val Gin Ala Ser Ser Gin Cys Phe Gin Ser 1685 1690 1695 -133- Ala Thr Asp Val Ala Ala Phe Leu Gly Ala Leu Ala Ser Leu Gly Ser 1700 1705 1710 Leu Asn He Pro Tyr Lys He Glu Ala Val Gin Ser Glu Thr Val Glu 1715 1720 1725 Pro Pro Pro Pro Ala Gin Leu His Phe Met Tyr Val Ala Ala Ala Ala 1730 1735 1740 Phe Val Leu Leu Phe Phe Val Gly Cys Gly Val Leu Leu Ser Arg Lys 1745 1750 1755 1760 Arg Arg Arg Gin His Gly Gin Leu Trp Phe Pro Glu Gly Phe Lys Val 1765 1770 1775 Ser Glu Ala Ser Lys Lys Lys Arg Arg Glu Glu Leu Gly Glu Asp Ser 1780 1785 1790 Val Gly Leu Lys Pro Leu Lys Asn Ala Ser Asp Gly Ala Leu Met Asp 1795 1800 1805 Asp Asn Gin Asn Glu Trp Gly Asp Glu Asp Leu Glu Thr Lys Lys Phe 1810 1815 1820 Arg Phe Glu Glu Pro Val Val Leu Pro Asp Leu Asp Asp Gin Thr Asp 1825 1830 1835 1840 His Arg Gin Trp Thr Gin Gin His Leu Asp Ala Ala Asp Leu Arg Met 1845 1850 1855 Ser Ala Met Ala Pro Thr Pro Pro Gin Gly Glu Val Asp Ala Asp Cys 1860 1865 1870 Met Asp Val Asn Val Arg Gly Pro Asp Gly Phe Thr Pro Leu Met He 1875 1880 1885 Ala Ser Cys Ser Gly Gly Gly Leu Glu Thr Gly Asn Ser Glu Glu Glu 1890 1895 1900 Glu Asp Ala Pro Ala Val He Ser Asp Phe He Tyr Gin Gly Ala Ser 1905 1910 1915 1920 Leu His Asn Gin Thr Asp Arg Thr Gly Glu Thr Ala Leu His Leu Ala 1925 1930 1935 Ala Arg Tyr Ser Arg Ser Asp Ala Ala Lys Arg Leu Leu Glu Ala Ser 1940 1945 1950 Ala Asp Ala Asn He Gin Asp Asn Met Gly Arg Thr Pro Leu His Ala 1955 1960 1965 Ala Val Ser Ala Asp Ala Gin Gly Val Phe Gin He Leu He Arg Asn 1970 1975 1980 Arg Ala Thr Asp Leu Asp Ala Arg Met His Asp Gly Thr Thr Pro Leu 1985 1990 1995 2000 He Leu Ala Ala Arg Leu Ala Val Glu Gly Met Leu Glu Asp Leu He 2005 2010 2015 Asn Ser His Ala Asp Val Asn Ala Val Asp Asp Leu Gly Lys Ser Ala 2020 2025 2030 Leu His Trp Ala Ala Ala Val Asn Asn Val Asp Ala Ala Val Val Leu 2035 2040 2045 Leu Lys Asn Gly Ala Asn Lys Asp Met Gin Asn Asn Arg Glu Glu Thr -134- 2050 2055 2060 Pro Leu Phe Leu Ala Ala Arg Glu Gly Ser Tyr Glu Thr Ala Lys Val 2065 2070 2075 2080 Leu Leu Asp His Phe Ala Asn Arg Asp He Thr Asp His Met Asp Arg 2085 2090 2095 Leu Pro Arg Asp He Ala Gin Glu Arg Met His His Asp He Val Arg 2100 2105 2110 Leu Leu Asp Glu Tyr Asn Leu Val Arg Ser Pro Gin Leu His Gly Ala 2115 2120 2125 Pro Leu Gly Gly Thr Pro Thr Leu Ser Pro Pro Leu Cys Ser Pro Asn 2130 2135 2140 Gly Tyr Leu Gly Ser Leu Lys Pro Gly Val Gin Gly Lys Lys Val Arg 2145 2150 2155 2160 Lys Pro Ser Ser Lys Gly Leu Ala Cys Gly Ser Lys Glu Ala Lys Asp 2165 2170 2175 Leu Lys Ala Arg Arg Lys Lys Ser Gin Asp Gly Lys Gly Cys Leu Leu 2180 2185 2190 Asp Ser Ser Gly Met Leu Ser Pro Val Asp Ser Leu Glu Ser Pro His 2195 2200 2205 Gly Tyr Leu Ser Asp Val Ala Ser Pro Pro Leu Leu Pro Ser Pro Phe 2210 2215 2220 Gin Gin Ser Pro Ser Val Pro Leu Asn His Leu Pro Gly Met Pro Asp 2225 2230 2235 2240 Thr His Leu Gly He Gly His Leu Asn Val Ala Ala Lys Pro Glu Met 2245 2250 2255 Ala Ala Leu Gly Gly Gly Gly Arg Leu Ala Phe Glu Thr Gly Pro Pro 2260 2265 2270 Arg Leu Ser His Leu Pro Val Ala Ser Gly Thr Ser Thr Val Leu Gly 2275 2280 2285 Ser Ser Ser Gly Gly Ala Leu Asn Phe Thr Val Gly Gly Ser Thr Ser 2290 2295 2300 Leu Asn Gly Gin Cys Glu Trp Leu Ser Arg Leu Gin Ser Gly Met Val 2305 2310 2315 2320 Pro Asn Gin Tyr Asn Pro Leu Arg Gly Ser Val Ala Pro Gly Pro Leu 2325 2330 2335 Ser Thr Gin Ala Pro Ser Leu Gin His Gly Met Val Gly Pro Leu His 2340 2345 2350 Ser Ser Leu Ala Ala Ser Ala Leu Ser Gin Met Met Ser Tyr Gin Gly 2355 2360 2365 Leu Pro Ser Thr Arg Leu Ala Thr Gin Pro His Leu Val Gin Thr Gin 2370 2375 2380 Gin Val Gin Pro Gin Asn Leu Gin Met Gin Gin Gin Asn Leu Gin Pro 2385 2390 2395 2400 Ala Asn He Gin Gin Gin Gin Ser Leu Gin Pro Pro Pro Pro Pro Pro 2405 2410 2415 -135- Gln Pro His Leu Gly Val Ser Ser Ala Ala Ser Gly His Leu Gly Arg 2420 2425 2430 Ser Phe Leu Ser Gly Glu Pro Ser Gin Ala Asp Val Gin Pro Leu Gly 2435 2440 2445 Pro Ser Ser Leu Ala Val His Thr He Leu Pro Gin Glu Ser Pro Ala 2450 2455 2460 Leu Pro Thr Ser Leu Pro Ser Ser Leu Val Pro Pro Val Thr Ala Ala 2465 2470 2475 2480 Gin Phe Leu Thr Pro Pro Ser Gin His Ser Tyr Ser Ser Pro Val Glu 2485 2490 2495 Asn Thr Pro Ser His Gin Leu Gin Val Pro Glu His Pro Phe Leu Thr 2500 2505 2510 Pro Ser Pro Glu Ser Pro Asp Gin Trp Ser Ser Ser Ser Pro His Ser 2515 2520 2525 Asn Val Ser Asp Trp Ser Glu Gly Val Ser Ser Pro Pro Thr Ser Met 2530 2535 2540 Gin Ser Gin He Ala Arg He Pro Glu Ala Phe Lys 2545 2550 2555 (2) INFORMATION FOR SEQ ID NO:21: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9723 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: cDNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 10..7419 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: GGAATTCCG CCC GCC CTG CGC CCC GCT CTG CTG TGG GCG CTG CTG GCG 48 Pro Ala Leu Arg Pro Ala Leu Leu Trp Ala Leu Leu Ala 1 5 10 CTC TGG CTG TGC TGC GCG GCC CCC GCG CAT GCA TTG CAG TGT CGA GAT 96 Leu Trp Leu Cys Cys Ala Ala Pro Ala His Ala Leu Gin Cys Arg Asp 15 20 25 GGC TAT GAA CCC TGT GTA AAT GAA GGA ATG TGT GTT ACC TAC CAC AAT 144 Gly Tyr Glu Pro Cys Val Asn Glu Gly Met Cys Val Thr Tyr His Asn 30 35 40 45 GGC ACA GGA TAC TGC AAA TGT CCA GAA GGC TTC TTG GGG GAA TAT TGT 192 Gly Thr Gly Tyr Cys Lys Cys Pro Glu Gly Phe Leu Gly Glu Tyr Cys . 50 55 60 CAA CAT CGA GAC CCC TGT GAG AAG AAC CGC TGC CAG AAT GGT GGG ACT 240 Gin His Arg Asp Pro Cys Glu Lys Asn Arg Cys Gin Asn Gly Gly Thr 65 70 75 TGT GTG GCC CAG GCC ATG CTG GGG AAA GCC ACG TGC CGA TGT GCC TCA 288 Cys Val Ala Gin Ala Met Leu Gly Lys Ala Thr Cys Arg Cys Ala Ser -136- 80 85 90 GGG TTT ACA GGA GAG GAC TGC CAG TAC TCA ACA TCT CAT CCA TGC TTT 336 Gly Phe Thr Gly Glu Asp Cys Gin Tyr Ser Thr Ser His Pro Cys Phe 95 100 105 GTG TCT CGA CCC TGC CTG AAT GGC GGC ACA TGC CAT ATG CTC AGC CGG 384 Val Ser Arg Pro Cys Leu Asn Gly Gly Thr Cys His Met Leu Ser Arg 110 115 120 125 GAT ACC TAT GAG TGC ACC TGT CAA GTC GGG TTT ACA GGT AAG GAG TGC 432 Asp Thr Tyr Glu Cys Thr Cys Gin Val Gly Phe Thr Gly Lys Glu Cys 130 135 140 CAA TGG ACG GAT GCC TGC CTG TCT CAT CCC TGT GCA AAT GGA AGT ACC 480 Gin Trp Thr Asp Ala Cys Leu Ser His Pro Cys Ala Asn Gly Ser Thr 145 150 155 TGT ACC ACT GTG GCC AAC CAG TTC TCC TGC AAA TGC CTC ACA GGC TTC 528 Cys Thr Thr Val Ala Asn Gin Phe Ser Cys Lys Cys Leu Thr Gly Phe 160 165 170 ACA GGG CAG AAA TGT GAG ACT GAT GTC AAT GAG TGT GAC ATT CCA GGA 576 Thr Gly Gin Lys Cys Glu Thr Asp Val Asn Glu Cys Asp He Pro Gly 175 180 185 CAC TGC CAG CAT GGT GGC ACC TGC CTC AAC CTG CCT GGT TCC TAC CAG 624 His Cys Gin His Gly Gly Thr Cys Leu Asn Leu Pro Gly Ser Tyr Gin 190 195 200 205 TGC CAG TGC CCT CAG GGC TTC ACA GGC CAG TAC TGT GAC AGC CTG TAT 672 Cys Gin Cys Pro Gin Gly Phe Thr Gly Gin Tyr Cys Asp Ser Leu Tyr 210 215 220 GTG CCC TGT GCA CCC TCA CCT TGT GTC AAT GGA GGC ACC TGT CGG CAG 720 Val Pro Cys Ala Pro Ser Pro Cys Val Asn Gly Gly Thr Cys Arg Gin 225 230 235 ACT GGT GAC TTC ACT TTT GAG TGC AAC TGC CTT CCA GGT TTT GAA GGG 768 Thr Gly Asp Phe Thr Phe Glu Cys Asn Cys Leu Pro Gly Phe Glu Gly 240 245 250 AGC ACC TGT GAG AGG AAT ATT GAT GAC TGC CCT AAC CAC AGG TGT CAG 816 Ser Thr Cys Glu Arg Asn He Asp Asp Cys Pro Asn His Arg Cys Gin 255 260 265 AAT GGA GGG GTT TGT GTG GAT GGG GTC AAC ACT TAC AAC TGC CGC TGT 864 Asn Gly Gly Val Cys Val Asp Gly Val Asn Thr Tyr Asn Cys Arg Cys 270 275 280 285 CCC CCA CAA TGG ACA GGA CAG TTC TGC ACA GAG GAT GTG GAT GAA TGC 912 Pro Pro Gin Trp Thr Gly Gin Phe Cys Thr Glu Asp Val Asp Glu Cys 290 295 300 CTG CTG CAG CCC AAT GCC TGT CAA AAT GGG GGC ACC TGT GCC AAC CGC 960 Leu Leu Gin Pro Asn Ala Cys Gin Asn Gly Gly Thr Cys Ala Asn Arg 305 310 315 AAT GGA GGC TAT GGC TGT GTA TGT GTC AAC GGC TGG AGT GGA GAT GAC 1008 Asn Gly Gly Tyr Gly Cys Val Cys Val Asn Gly Trp Ser Gly Asp Asp 320 325 330 TGC AGT GAG AAC ATT GAT GAT TGT GCC TTC GCC TCC TGT ACT CCA GGC 1056 Cys Ser Glu Asn He Asp Asp Cys Ala Phe Ala Ser Cys Thr Pro Gly 335 340 345 TCC ACC TGC ATC GAC CGT GTG GCC TCC TTC TCT TGC ATG TGC CCA GAG 1104 -137- Ser Thr Cys He Asp Arg Val Ala Ser Phe Ser Cys Met Cys Pro Glu 350 355 360 365 GGG AAG GCA GGT CTC CTG TGT CAT CTG GAT GAT GCA TGC ATC AGC AAT 1152 Gly Lys Ala Gly Leu Leu Cys His Leu Asp Asp Ala Cys He Ser Asn 370 375 380 CCT TGC CAC AAG GGG GCA CTG TGT GAC ACC AAC CCC CTA AAT GGG CAA 1200 Pro Cys His Lys Gly Ala Leu Cys Asp Thr Asn Pro Leu Asn Gly Gin 385 390 395 TAT ATT TGC ACC TGC CCA CAA GGC TAC AAA GGG GCT GAC TGC ACA GAA 1248 Tyr He Cys Thr Cys Pro Gin Gly Tyr Lys Gly Ala Asp Cys Thr Glu 400 405 410 GAT GTG GAT GAA TGT GCC ATG GCC AAT AGC AAT CCT TGT GAG CAT GCA 1296 Asp Val Asp Glu Cys Ala Met Ala Asn Ser Asn Pro Cys Glu His Ala 415 420 425 GGA AAA TGT GTG AAC ACG GAT GGC GCC TTC CAC TGT GAG TGT CTG AAG 1344 Gly Lys Cys Val Asn Thr Asp Gly Ala Phe His Cys Glu Cys Leu Lys 430 435 440 445 GGT TAT GCA GGA CCT CGT TGT GAG ATG GAC ATC AAT GAG TGC CAT TCA 1392 Gly Tyr Ala Gly Pro Arg Cys Glu Met Asp He Asn Glu Cys His Ser 450 455 460 GAC CCC TGC CAG AAT GAT GCT ACC TGT CTG GAT AAG ATT GGA GGC TTC 1440 Asp Pro Cys Gin Asn Asp Ala Thr Cys Leu Asp Lys He Gly Gly Phe 465 470 475 ACA TGT CTG TGC ATG CCA GGT TTC AAA GGT GTG CAT TGT GAA TTA GAA 1488 Thr Cys Leu Cys Met Pro Gly Phe Lys Gly Val His Cys Glu Leu Glu 480 485 490 ATA AAT GAA TGT CAG AGC AAC CCT TGT GTG AAC AAT GGG CAG TGT GTG 1536 He Asn Glu Cys Gin Ser Asn Pro Cys Val Asn Asn Gly Gin Cys Val 495 500 505 GAT AAA GTC AAT CGT TTC CAG TGC CTG TGT CCT CCT GGT TTC ACT GGG 1584 Asp Lys Val Asn Arg Phe Gin Cys Leu Cys Pro Pro Gly Phe Thr Gly 510 515 520 525 CCA GTT TGC CAG ATT GAT ATT GAT GAC TGT TCC AGT ACT CCG TGT CTG 1632 Pro Val Cys Gin He Asp He Asp Asp Cys Ser Ser Thr Pro Cys Leu 530 535 540 AAT GGG GCA AAG TGT ATC GAT CAC CCG AAT GGC TAT GAA TGC CAG TGT 1680 Asn Gly Ala Lys Cys He Asp His Pro Asn Gly Tyr Glu Cys Gin Cys 545 550 555 GCC ACA GGT TTC ACT GGT GTG TTG TGT GAG GAG AAC ATT GAC AAC TGT 1728 Ala Thr Gly Phe Thr Gly Val Leu Cys Glu Glu Asn He Asp Asn Cys 560 565 570 GAC CCC GAT CCT TGC CAC CAT GGT CAG TGT CAG GAT GGT ATT GAT TCC 1776 Asp Pro Asp Pro Cys His His Gly Gin Cys Gin Asp Gly He Asp Ser 575 580 585 TAC ACC TGC ATC TGC AAT CCC GGG TAC ATG GGC GCC ATC TGC AGT GAC 1824 Tyr Thr Cys He Cys Asn Pro Gly Tyr Met Gly Ala He Cys Ser Asp 590 595 600 605 CAG ATT GAT GAA TGT TAC AGC AGC CCT TGC CTG AAC GAT GGT CGC TGC 1872 Gin He Asp Glu Cys Tyr Ser Ser Pro Cys Leu Asn Asp Gly Arg Cys 610 615 620 -138- ATT GAC CTG GTC AAT GGC TAC CAG TGC AAC TGC CAG CCA GGC ACG TCA 1920 He Asp Leu Val Asn Gly Tyr Gin Cys Asn Cys Gin Pro Gly Thr Ser 625 630 635 GGG GTT AAT TGT GAA ATT AAT TTT GAT GAC TGT GCA AGT AAC CCT TGT 1968 Gly Val Asn Cys Glu He Asn Phe Asp Asp Cys Ala Ser Asn Pro Cys 640 645 650 ATC CAT GGA ATC TGT ATG GAT GGC ATT AAT CGC TAC AGT TGT GTC TGC 2016 He His Gly He Cys Met Asp Gly He Asn Arg Tyr Ser Cys Val Cys 655 660 665 TCA CCA GGA TTC ACA GGG CAG AGA TGT AAC ATT GAC ATT GAT GAG TGT 2064 Ser Pro Gly Phe Thr Gly Gin Arg Cys Asn He Asp He Asp Glu Cys 670 675 680 685 GCC TCC AAT CCC TGT CGC AAG GGT GCA ACA TGT ATC AAC GGT GTG AAT 2112 Ala Ser Asn Pro Cys Arg Lys Gly Ala Thr Cys He Asn Gly Val Asn 690 695 700 GGT TTC CGC TGT ATA TGC CCC GAG GGA CCC CAT CAC CCC AGC TGC TAC 2160 Gly Phe Arg Cys He Cys Pro Glu Gly Pro His His Pro Ser Cys Tyr 705 710 715 TCA CAG GTG AAC GAA TGC CTG AGC AAT CCC TGC ATC CAT GGA AAC TGT 2208 Ser Gin Val Asn Glu Cys Leu Ser Asn Pro Cys He His Gly Asn Cys 720 725 730 ACT GGA GGT CTC AGT GGA TAT AAG TGT CTC TGT GAT GCA GGC TGG GTT 2256 Thr Gly Gly Leu Ser Gly Tyr Lys Cys Leu Cys Asp Ala Gly Trp Val 735 740 745 GGC ATC AAC TGT GAA GTG GAC AAA AAT GAA TGC CTT TCG AAT CCA TGC 2304 Gly He Asn Cys Glu Val Asp Lys Asn Glu Cys Leu Ser Asn Pro Cys 750 755 760 765 CAG AAT GGA GGA ACT TGT GAC AAT CTG GTG AAT GGA TAC AGG TGT ACT 2352 Gin Asn Gly Gly Thr Cys, Asp Asn Leu Val Asn Gly Tyr Arg Cys Thr 770 775 780 TGC AAG AAG GGC TTT AAA GGC TAT AAC TGC CAG GTG AAT ATT GAT GAA 2400 Cys Lys Lys Gly Phe Lys Gly Tyr Asn Cys Gin Val Asn He Asp Glu 785 790 795 TGT GCC TCA AAT CCA TGC CTG AAC CAA GGA ACC TGC TTT GAT GAC ATA 2448 Cys Ala Ser Asn Pro Cys Leu Asn Gin Gly Thr Cys Phe Asp Asp He 800 805 810 AGT GGC TAC ACT TGC CAC TGT GTG CTG CCA TAC ACA GGC AAG AAT TGT 2496 Ser Gly Tyr Thr Cys His Cys Val Leu Pro Tyr Thr Gly Lys Asn Cys 815 820 825 CAG ACA GTA TTG GCT CCC TGT TCC CCA AAC CCT TGT GAG AAT GCT GCT 2544 Gin Thr Val Leu Ala Pro Cys Ser Pro Asn Pro Cys Glu Asn Ala Ala 830 835 840 845 GTT TGC AAA GAG TCA CCA AAT TTT GAG AGT TAT ACT TGC TTG TGT GCT 2592 Val Cys Lys Glu Ser Pro Asn Phe Glu Ser Tyr Thr Cys Leu Cys Ala 850 855 860 CCT GGC TGG CAA GGT CAG CGG TGT ACC ATT GAC ATT GAC GAG TGT ATC 2640 Pro Gly Trp Gin Gly Gin Arg Cys Thr He Asp He Asp Glu Cys He 865 870 875 TCC AAG CCC TGC ATG AAC CAT GGT CTC TGC CAT AAC ACC CAG GGC AGC 2688 Ser Lys Pro Cys Met Asn His Gly Leu Cys His Asn Thr Gin Gly Ser 880 885 890 -139- TAC ATG TGT GAA TGT CCA CCA GGC TTC AGT GGT ATG GAC TGT GAG GAG 2736 Tyr Met Cys Glu Cys Pro Pro Gly Phe Ser Gly Met Asp Cys Glu Glu 895 900 905 GAC ATT GAT GAC TGC CTT GCC AAT CCT TGC CAG AAT GGA GGT TCC TGT 2784 Asp He Asp Asp Cys Leu Ala Asn Pro Cys Gin Asn Gly Gly Ser Cys 910 915 920 925 ATG GAT GGA GTG AAT ACT TTC TCC TGC CTC TGC CTT CCG GGT TTC ACT 2832 Met Asp Gly Val Asn Thr Phe Ser Cys Leu Cys Leu Pro Gly Phe Thr 930 935 940 GGG GAT AAG TGC CAG ACA GAC ATG AAT GAG TGT CTG AGT GAA CCC TGT 2880 Gly Asp Lys Cys Gin Thr Asp Met Asn Glu Cys Leu Ser Glu Pro Cys 945 950 955 AAG AAT GGA GGG ACC TGC TCT GAC TAC GTC AAC AGT TAC ACT TGC AAG 2928 Lys Asn Gly Gly Thr Cys Ser Asp Tyr Val Asn Ser Tyr Thr Cys Lys 960 965 970 TGC CAG GCA GGA TTT GAT GGA GTC CAT TGT GAG AAC AAC ATC AAT GAG 2976 Cys Gin Ala Gly Phe Asp Gly Val His Cys Glu Asn Asn He Asn Glu 975 980 985 TGC ACT GAG AGC TCC TGT TTC AAT GGT GGC ACA TGT GTT GAT GGG ATT 3024 Cys Thr Glu Ser Ser Cys Phe Asn Gly Gly Thr Cys Val Asp Gly He 990 995 1000 1005 AAC TCC TTC TCT TGC TTG TGC CCT GTG GGT TTC ACT GGA TCC TTC TGC 3072 Asn Ser Phe Ser Cys Leu Cys Pro Val Gly Phe Thr Gly Ser Phe Cys 1010 1015 1020 CTC CAT GAG ATC AAT GAA TGC AGC TCT CAT CCA TGC CTG AAT GAG GGA 3120 Leu His Glu He Asn Glu Cys Ser Ser His Pro Cys Leu Asn Glu Gly 1025 1030 1035 ACG TGT GTT GAT GGC CTG GGT ACC TAC CGC TGC AGC TGC CCC CTG GGC 3168 Thr Cys Val Asp Gly Leu Gly Thr Tyr Arg Cys Ser Cys Pro Leu Gly 1040 1045 1050 TAC ACT GGG AAA AAC TGT CAG ACC CTG GTG AAT CTC TGC AGT CGG TCT 3216 Tyr Thr Gly Lys Asn Cys Gin Thr Leu Val Asn Leu Cys Ser Arg Ser 1055 1060 1065 CCA TGT AAA AAC AAA GGT ACT TGT GTT CAG AAA AAA GCA GAG TCC CAG 3264 Pro Cys Lys Asn Lys Gly Thr Cys Val Gin Lys Lys Ala Glu Ser Gin 1070 1075 1080 1085 TGC CTA TGT CCA TCT GGA TGG GCT GGT GCC TAT TGT GAC GTG CCC AAT 3312 Cys Leu Cys Pro Ser Gly Trp Ala Gly Ala Tyr Cys Asp Val Pro Asn 1090 1095 1100 GTC TCT TGT GAC ATA GCA GCC TCC AGG AGA GGT GTG CTT GTT GAA CAC 3360 Val Ser Cys Asp He Ala Ala Ser Arg Arg Gly Val Leu Val Glu His 1105 1110 1115 TTG TGC CAG CAC TCA GGT GTC TGC ATC AAT GCT GGC AAC ACG CAT TAC 3408 Leu Cys Gin His Ser Gly Val Cys He Asn Ala Gly Asn Thr His Tyr 1120 1125 1130 TGT CAG TGC CCC CTG GGC TAT ACT GGG AGC TAC TGT GAG GAG CAA CTC 3456 Cys Gin Cys Pro Leu Gly Tyr Thr Gly Ser Tyr Cys Glu Glu Gin Leu 1135 1140 1145 GAT GAG TGT GCG TCC AAC CCC TGC CAG CAC GGG GCA ACA TGC AGT GAC 3504 Asp Glu Cys Ala Ser Asn Pro Cys Gin His Gly Ala Thr Cys Ser Asp 1150 1155 1160 1165 -140- TTC ATT GGT GGA TAC AGA TGC GAG TGT GTC CCA GGC TAT CAG GGT GTC 3552 Phe He Gly Gly Tyr Arg Cys Glu Cys Val Pro Gly Tyr Gin Gly Val 1170 1175 1180 AAC TGT GAG TAT GAA GTG GAT GAG TGC CAG AAT CAG CCC TGC CAG AAT 3600 Asn Cys Glu Tyr Glu Val Asp Glu Cys Gin Asn Gin Pro Cys Gin Asn 1185 1190 1195 GGA GGC ACC TGT ATT GAC CTT GTG AAC CAT TTC AAG TGC TCT TGC CCA 3648 Gly Gly Thr Cys He Asp Leu Val Asn His Phe Lys Cys Ser Cys Pro 1200 1205 1210 CCA GGC ACT CGG GGC CTA CTC TGT GAA GAG AAC ATT GAT GAC TGT GCC 3696 Pro Gly Thr Arg Gly Leu Leu Cys Glu Glu Asn He Asp Asp Cys Ala 1215 1220 1225 CGG GGT CCC CAT TGC CTT AAT GGT GGT CAG TGC ATG GAT AGG ATT GGA 3744 Arg Gly Pro His Cys Leu Asn Gly Gly Gin Cys Met Asp Arg He Gly 1230 1235 1240 1245 GGC TAC AGT TGT CGC TGC TTG CCT GGC TTT GCT GGG GAG CGT TGT GAG 3792 Gly Tyr Ser Cys Arg Cys Leu Pro Gly Phe Ala Gly Glu Arg Cys Glu 1250 1255 1260 GGA GAC ATC AAC GAG TGC CTC TCC AAC CCC TGC AGC TCT GAG GGC AGC 3840 Gly Asp He Asn Glu Cys Leu Ser Asn Pro Cys Ser Ser Glu Gly Ser 1265 1270 1275 CTG GAC TGT ATA CAG CTC ACC AAT GAC TAC CTG TGT GTT TGC CGT AGT 3888 Leu Asp Cys He Gin Leu Thr Asn Asp Tyr Leu Cys Val Cys Arg Ser 1280 1285 1290 GCC TTT ACT GGC CGG CAC TGT GAA ACC TTC GTC GAT GTG TGT CCC CAG 3936 Ala Phe Thr Gly Arg His Cys Glu Thr Phe Val Asp Val Cys Pro Gin 1295 1300 1305 ATG CCC TGC CTG AAT GGA GGG ACT TGT GCT GTG GCC AGT AAC ATG CCT 3984 Met Pro Cys Leu Asn Gly Gly Thr Cys Ala Val Ala Ser Asn Met Pro 1310 1315 1320 1325 GA1 GGT TTC ATT TGC CGT TGT CCC CCG GGA TTT TCC GGG GCA AGG TGC 4032 Asp Gly Phe He Cys Arg Cys Pro Pro Gly Phe Ser Gly Ala Arg Cys 1330 1335 1340 CAG AGC AGC TGT GGA CAA GTG AAA TGT AGG AAG GGG GAG CAG TGT GTG 4080 Gin Ser Ser Cys Gly Gin Val Lys Cys Arg Lys Gly Glu Gin Cys Val 1345 1350 1355 CAC ACC GCC TCT GGA CCC CGC TGC TTC TGC CCC AGT CCC CGG GAC TGC 4128 His Thr Ala Ser Gly Pro Arg Cys Phe Cys Pro Ser Pro Arg Asp Cys 1360 1365 1370 GAG TCA GGC TGT GCC AGT AGC CCC TGC CAG CAC GGG GGC AGC TGC CAC 4176 Glu Ser Gly Cys Ala Ser Ser Pro Cys Gin His Gly Gly Ser Cys His 1375 1380 1385 CCT CAG CGC CAG CCT CCT TAT TAC TCC TGC CAG TGT GCC CCA CCA TTC 4224 Pro Gin Arg Gin Pro Pro Tyr Tyr Ser Cys Gin Cys Ala Pro Pro Phe 1390 1395 1400 1405 TCG GGT AGC CGC TGT GAA CTC TAC ACG GCA CCC CCC AGC ACC CCT CCT 4272 Ser Gly Ser Arg Cys Glu Leu Tyr Thr Ala Pro Pro Ser Thr Pro Pro 1410 1415 1420 GCC ACC TGT CTG AGC CAG TAT TGT GCC GAC AAA GCT CGG GAT GGC GTC 4320 Ala Thr Cys Leu Ser Gin Tyr Cys Ala Asp Lys Ala Arg Asp Gly Val 1425 1430 1435 -141- TGT GAT GAG GCC TGC AAC AGC CAT GCC TGC CAG TGG GAT GGG GGT GAC 4368 Cys Asp Glu Ala Cys Asn Ser His Ala Cys Gin Trp Asp Gly Gly Asp 1440 1445 1450 TGT TCT CTC ACC ATG GAG AAC CCC TGG GCC AAC TGC TCC TCC CCA CTT 4416 Cys Ser Leu Thr Met Glu Asn Pro Trp Ala Asn Cys Ser Ser Pro Leu 1455 1460 1465 CCC TGC TGG GAT TAT ATC AAC AAC CAG TGT GAT GAG CTG TGC AAC ACG 4464 Pro Cys Trp Asp Tyr He Asn Asn Gin Cys Asp Glu Leu Cys Asn Thr 1470 1475 1480 1485 GTC GAG TGC CTG TTT GAC AAC TTT GAA TGC CAG GGG AAC AGC AAG ACA 4512 Val Glu Cys Leu Phe Asp Asn Phe Glu Cys Gin Gly Asn Ser Lys Thr 1490 1495 1500 TGC AAG TAT GAC AAA TAC TGT GCA GAC CAC TTC AAA GAC AAC CAC TGT 4560 Cys Lys Tyr Asp Lys Tyr Cys Ala Asp His Phe Lys Asp Asn His Cys 1505 1510 1515 AAC CAG GGG TGC AAC AGT GAG GAG TGT GGT TGG GAT GGG CTG GAC TGT 4608 Asn Gin Gly Cys Asn Ser Glu Glu Cys Gly Trp Asp Gly Leu Asp Cys 1520 1525 1530 GCT GCT GAC CAA CCT GAG AAC CTG GCA GAA GGT ACC CTG GTT ATT GTG 4656 Ala Ala Asp Gin Pro Glu Asn Leu Ala Glu Gly Thr Leu Val He Val 1535 1540 1545 GTA TTG ATG CCA CCT GAA CAA CTG CTC CAG GAT GCT CGC AGC TTC TTG 4704 Val Leu Met Pro Pro Glu Gin Leu Leu Gin Asp Ala Arg Ser Phe Leu 1550 1555 1560 1565 CGG GCA CTG GGT ACC CTG CTC CAC ACC AAC CTG CGC ATT AAG CGG GAC 4752 Arg Ala Leu Gly Thr Leu Leu His Thr Asn Leu Arg He Lys Arg Asp 1570 1575 1580 TCC CAG GGG GAA CTC ATG GTG TAC CCC TAT TAT GGT GAG AAG TCA GCT 4800 Ser Gin Gly Glu Leu Met Val Tyr Pro Tyr Tyr Gly Glu Lys Ser Ala 1585 1590 1595 GCT ATG AAG AAA CAG AGG ATG ACA CGC AGA TCC CTT CCT GGT GAA CAA 4848 Ala Met Lys Lys Gin Arg Met Thr Arg Arg Ser Leu Pro Gly Glu Gin 1600 1605 1610 GAA CAG GAG GTG GCT GGC TCT AAA GTC TTT CTG GAA ATT GAC AAC CGC 4896 Glu Gin Glu Val Ala Gly Ser Lys Val Phe Leu Glu He Asp Asn Arg 1615 1620 1625 CAG TGT GTT CAA GAC TCA GAC CAC TGC TTC AAG AAC ACG GAT GCA GCA 4944 Gin Cys Val Gin Asp Ser Asp His Cys Phe Lys Asn Thr Asp Ala Ala 1630 1635 1640 1645 GCA GCT CTC CTG GCC TCT CAC GCC ATA CAG GGG ACC CTG TCA TAC CCT 4992 Ala Ala Leu Leu Ala Ser His Ala He Gin Gly Thr Leu Ser Tyr Pro 1650 1655 1660 CTT GTG TCT GTC GTC AGT GAA TCC CTG ACT CCA GAA CGC ACT CAG CTC 5040 Leu Val Ser Val Val Ser Glu Ser Leu Thr Pro Glu Arg Thr Gin Leu 1665 1670 1675 CTC TAT CTC CTT GCT GTT GCT GTT GTC ATC ATT CTG TTT ATT ATT CTG 5088 Leu Tyr Leu Leu Ala Val Ala Val Val He He Leu Phe He He Leu 1680 1685 1690 CTG GGG GTA ATC ATG GCA AAA CGA AAG CGT AAG CAT GGC TCT CTC TGG 5136 Leu Gly Val He Met Ala Lys Arg Lys Arg Lys His Gly Ser Leu Trp 1695 1700 1705 -142- CTG CCT GAA GGT TTC ACT CTT CGC CGA GAT GCA AGC AAT CAC AAG CGT 5184 Leu Pro Glu Gly Phe Thr Leu Arg Arg Asp Ala Ser Asn His Lys Arg 1710 1715 1720 1725 CGT GAG CCA GTG GGA CAG GAT GCT GTG GGG CTG AAA AAT CTC TCA GTG 5232 Arg Glu Pro Val Gly Gin Asp Ala Val Gly Leu Lys Asn Leu Ser Val 1730 1735 1740 CAA GTC TCA GAA GCT AAC CTA ATT GGT ACT GGA ACA AGT GAA CAC TGG 5280 Gin Val Ser Glu Ala Asn Leu He Gly Thr Gly Thr Ser Glu His Trp 1745 1750 1755 GTC GAT GAT GAA GGG CCC CAG CCA AAG AAA GTA AAG GCT GAA GAT GAG 5328 Val Asp Asp Glu Gly Pro Gin Pro Lys Lys Val Lys Ala Glu Asp Glu 1760 1765 1770 GCC TTA CTC TCA GAA GAA GAT GAC CCC ATT GAT CGA CGG CCA TGG ACA 5376 Ala Leu Leu Ser Glu Glu Asp Asp Pro He Asp Arg Arg Pro Trp Thr 1775 1780 1785 CAG CAG CAC CTT GAA GCT GCA GAC ATC CGT AGG ACA CCA TCG CTG GCT 5424 Gin Gin His Leu Glu Ala Ala Asp He Arg Arg Thr Pro Ser Leu Ala 1790 1795 1800 1805 CTC ACC CCT CCT CAG GCA GAG CAG GAG GTG GAT GTG TTA GAT GTG AAT 5472 Leu Thr Pro Pro Gin Ala Glu Gin Glu Val Asp Val Leu Asp Val Asn 1810 1815 1820 GTC CGT GGC CCA GAT GGC TGC ACC CCA TTG ATG TTG GCT TCT CTC CGA 5520 Val Arg Gly Pro Asp Gly Cys Thr Pro Leu Met Leu Ala Ser Leu Arg 1825 1830 1835 GGA GGC AGC TCA GAT TTG AGT GAT GAA GAT GAA GAT GCA GAG GAC TCT 5568 Gly Gly Ser Ser Asp Leu Ser Asp Glu Asp Glu Asp Ala Glu Asp Ser 1840 1845 1850 TCT GCT AAC ATC ATC ACA GAC TTG GTC TAC CAG GGT GCC AGC CTC CAG 5616 Ser Ala Asn He He Thr Asp Leu Val Tyr Gin Gly Ala Ser Leu Gin 1855 1860 1865 GCC CAG ACA GAC CGG ACT GGT GAG ATG GCC CTG CAC CTT GCA GCC CGC 5664 Ala Gin Thr Asp Arg Thr Gly Glu Met Ala Leu His Leu Ala Ala Arg 1870 1875 1880 1885 TAC TCA CGG GCT GAT GCT GCC AAG CGT CTC CTG GAT GCA GGT GCA GAT 5712 Tyr Ser Arg Ala Asp Ala Ala Lys Arg Leu Leu Asp Ala Gly Ala Asp 1890 1895 1900 GCC AAT GCC CAG GAC AAC ATG GGC CGC TGT CCA CTC CAT GCT GCA GTG 5760 Ala Asn Ala Gin Asp Asn Met Gly Arg Cys Pro Leu His Ala Ala Val 1905 1910 1915 GCA GCT GAT GCC CAA GGT GTC TTC CAG ATT CTG ATT CGC AAC CGA GTA 5808 Ala Ala Asp Ala Gin Gly Val Phe Gin He Leu He Arg Asn Arg Val 1920 1925 1930 ACT GAT CTA GAT GCC AGG ATG AAT GAT GGT ACT ACA CCC CTG ATC CTG 5856 Thr Asp Leu Asp Ala Arg Met Asn Asp Gly Thr Thr Pro Leu He Leu 1935 1940 1945 GCT GCC CGC CTG GCT GTG GAG GGA ATG GTG GCA GAA CTG ATC AAC TGC 5904 Ala Ala Arg Leu Ala Val Glu Gly Met Val Ala Glu Leu He Asn Cys 1950 1955 1960 1965 CAA GCG GAT GTG AAT GCA GTG GAT GAC CAT GGA AAA TCT GCT CTT CAC 5952 Gin Ala Asp Val Asn Ala Val Asp Asp His Gly Lys Ser Ala Leu His 1970 1975 1980 -143- TGG GCA GCT GCT GTC AAT AAT GTG GAG GCA ACT CTT TTG TTG TTG AAA 6000 Trp Ala Ala Ala Val Asn Asn Val Glu Ala Thr Leu Leu Leu Leu Lys 1985 1990 1995 AAT GGG GCC AAC CGA GAC ATG CAG GAC AAC AAG GAA GAG ACA CCT CTG 6048 Asn Gly Ala Asn Arg Asp Met Gin Asp Asn Lys Glu Glu Thr Pro Leu 2000 2005 2010 TTT CTT GCT GCC CGG GAG GGG AGC TAT GAA GCA GCC AAG ATC CTG TTA 6096 Phe Leu Ala Ala Arg Glu Gly Ser Tyr Glu Ala Ala Lys He Leu Leu 2015 2020 2025 GAC CAT TTT GCC AAT CGA GAC ATC ACA GAC CAT ATG GAT CGT CTT CCC 6144 Asp His Phe Ala Asn Arg Asp He Thr Asp His Met Asp Arg Leu Pro 2030 2035 2040 2045 CGG GAT GTG GCT CGG GAT CGC ATG CAC CAT GAC ATT GTG CGC CTT CTG 6192 Arg Asp Val Ala Arg Asp Arg Met His His Asp He Val Arg Leu Leu 2050 2055 2060 GAT GAA TAC AAT GTG ACC CCA AGC CCT CCA GGC ACC GTG TTG ACT TCT 6240 Asp Glu Tyr Asn Val Thr Pro Ser Pro Pro Gly Thr Val Leu Thr Ser 2065 2070 2075 GCT CTC TCA CCT GTC ATC TGT GGG CCC AAC AGA TCT TTC CTC AGC CTG 6288 Ala Leu Ser Pro Val He Cys Gly Pro Asn Arg Ser Phe Leu Ser Leu 2080 2085 2090 AAG CAC ACC CCA ATG GGC AAG AAG TCT AGA CGG CCC AGT GCC AAG AGT 6336 Lys His Thr Pro Met Gly Lys Lys Ser Arg Arg Pro Ser Ala Lys Ser 2095 2100 2105 ACC ATG CCT ACT AGC CTC CCT AAC CTT GCC AAG GAG GCA AAG GAT GCC 6384 Thr Met Pro Thr Ser Leu Pro Asn Leu Ala Lys Glu Ala Lys Asp Ala 2110 2115 2120 2125 AAG GGT AGT AGG AGG AAG AAG TCT CTG AGT GAG AAG GTC CAA CTG TCT 6432 Lys Gly Ser Arg Arg Lys Lys Ser Leu Ser Glu Lys Val Gin Leu Ser 2130 2135 2140 GAG AGT TCA GTA ACT TTA TCC CCT GTT GAT TCC CTA GAA TCT CCT CAC 6480 Glu Ser Ser Val Thr Leu Ser Pro Val Asp Ser Leu Glu Ser Pro His 2145 2150 2155 ACG TAT GTT TCC GAC ACC ACA TCC TCT CCA ATG ATT ACA TCC CCT GGG 6528 Thr Tyr Val Ser Asp Thr Thr Ser Ser Pro Met He Thr Ser Pro Gly 2160 2165 2170 ATC TTA CAG GCC TCA CCC AAC CCT ATG TTG GCC ACT GCC GCC CCT CCT 6576 He Leu Gin Ala Ser Pro Asn Pro Met Leu Ala Thr Ala Ala Pro Pro 2175 2180 2185 GCC CCA GTC CAT GCC CAG CAT GCA CTA TCT TTT TCT AAC CTT CAT GAA 6624 Ala Pro Val His Ala Gin His Ala Leu Ser Phe Ser Asn Leu His Glu 2190 2195 2200 2205 ATG CAG CCT TTG GCA CAT GGG GCC AGC ACT GTG CTT CCC TCA GTG AGC 6672 Met Gin Pro Leu Ala His Gly Ala Ser Thr Val Leu Pro Ser Val Ser 2210 2215 2220 CAG TTG CTA TCC CAC CAC CAC ATT GTG TCT CCA GGC AGT GGC AGT GCT 6720 Gin Leu Leu Ser His His His He Val Ser Pro Gly Ser Gly Ser Ala 2225 2230 2235 GGA AGC TTG AGT AGG CTC CAT CCA GTC CCA GTC CCA GCA GAT TGG ATG 6768 Gly Ser Leu Ser Arg Leu His Pro Val Pro Val Pro Ala Asp Trp Met 2240 2245 2250 -144- AAC CGC ATG GAG GTG AAT GAG ACC CAG TAC AAT GAG ATG TTT GGT ATG 6816 Asn Arg Met Glu Val Asn Glu Thr Gin Tyr Asn Glu Met Phe Gly Met 2255 2260 2265 GTC CTG GCT CCA GCT GAG GGC ACC CAT CCT GGC ATA GCT CCC CAG AGC 6864 Val Leu Ala Pro Ala Glu Gly Thr His Pro Gly He Ala Pro Gin Ser 2270 2275 2280 2285 AGG CCA CCT GAA GGG AAG CAC ATA ACC ACC CCT CGG GAG CCC TTG CCC 6912 Arg Pro Pro Glu Gly Lys His He Thr Thr Pro Arg Glu Pro Leu Pro 2290 2295 2300 CCC ATT GTG ACT TTC CAG CTC ATC CCT AAA GGC AGT ATT GCC CAA CCA 6960 Pro He Val Thr Phe Gin Leu He Pro Lys Gly Ser He Ala Gin Pro 2305- 2310 2315 GCG GGG GCT CCC CAG CCT CAG TCC ACC TGC CCT CCA GCT GTT GCG GGC 7008 Ala Gly Ala Pro Gin Pro Gin Ser Thr Cys Pro Pro Ala Val Ala Gly 2320 2325 2330 CCC CTG CCC ACC ATG TAC CAG ATT CCA GAA ATG GCC CGT TTG CCC AGT 7056 Pro Leu Pro Thr Met Tyr Gin He Pro Glu Met Ala Arg Leu Pro Ser 2335 2340 2345 GTG GCT TTC CCC ACT GCC ATG ATG CCC CAG CAG GAC GGG CAG GTA GCT 7104 Val Ala Phe Pro Thr Ala Met Met Pro Gin Gin Asp Gly Gin Val Ala 2350 2355 2360 2365 CAG ACC ATT CTC CCA GCC TAT CAT CCT TTC CCA GCC TCT GTG GGC AAG 7152 Gin Thr He Leu Pro Ala Tyr His Pro Phe Pro Ala Ser Val Gly Lys 2370 2375 2380 TAC CCC ACA CCC CCT TCA CAG CAC AGT TAT GCT TCC TCA AAT GCT GCT 7200 Tyr Pro Thr Pro Pro Ser Gin His Ser Tyr Ala Ser Ser Asn Ala Ala 2385 2390 2395 GAG CGA ACA CCC AGT CAC AGT GGT CAC CTC CAG GGT GAG CAT CCC TAC 7248 Glu Arg Thr Pro Ser His Ser Gly His Leu Gin Gly Glu His Pro Tyr 2400 2405 2410 CTG ACA CCA TCC CCA GAG TCT CCT GAC CAG TGG TCA AGT TCA TCA CCC 7296 Leu Thr Pro Ser Pro Glu Ser Pro Asp Gin Trp Ser Ser Ser Ser Pro 2415 2420 2425 CAC TCT GCT TCT GAC TGG TCA JAT GTG ACC ACC AGC CCT ACC CCT GGG 7344 His Ser Ala Ser Asp Trp Ser Asp Val Thr Thr Ser Pro Thr Pro Gly 2430 2435 2440 2445 GGT GCT GGA GGA GGT CAG CGG GGA CCT GGG ACA CAC ATG TCT GAG CCA 7392 Gly Ala Gly Gly Gly Gin Arg Gly Pro Gly Thr His Met Ser Glu Pro 2450 2455 2460 CCA CAC AAC AAC ATG CAG GTT TAT GCG TGAGAGAGTC CACCTCCAGT 7439 Pro His Asn Asn Met Gin Val Tyr Ala 2465 2470 GTAGAGACAT AACTGACTTT TGTAAATGCT GCTGAGGAAC AAATGAAGGT CATCCGGGAG 7499 AGAAATGAAG AAATCTCTGG AGCCAGCTTC TAGAGGTAGG AAAGAGAAGA TGTTCTTATT 7559 CAGATAATGC AAGAGAAGCA ATTCGTCAGT TTCACTGGGT ATCTGCAAGG CTTATTGATT 7619 ATTCTAATCT AATAAGACAA GTTTGTGGAA ATGCAAGATG AATACAAGCC TTGGGTCCAT 7679 GTTTACTCTC TTCTATTTGG AGAATAAGAT GGATGCTTAT TGAAGCCCAG ACATTCTTGC 7739 AGCTTGGACT GCATTTTAAG CCCTGCAGGC TTCTGCCATA TCCATGAGAA GATTCTACAC 7799 -145- TAGCGTCCTG TTGGGAATTA TGCCCTGGAA TTCTGCCTGA ATTGACCTAC GCATCTCCTC 7859 CTCCTTGGAC ATTCTTTTGT CTTCATTTGG TGCTTTTGGT TTTGCACCTC TCCGTGATTG 7919 TAGCCCTACC AGCATGTTAT AGGGCAAGAC CTTTGTGCTT TTGATCATTC TGGCCCATGA 7979 AAGCAACTTT GGTCTCCTTT CCCCTCCTGT CTTCCCGGTA TCCCTTGGAG TCTCACAAGG 8039 TTTACTTTGG TATGGTTCTC AGCACAAACC TTTCAAGTAT GTTGTTTCTT TGGAAAATGG 8099 ACATACTGTA TTGTGTTCTC CTGCATATAT CATTCCTGGA GAGAGAAGGG GAGAAGAATA 8159 CTTTTCTTCA ACAAATTTTG GGGGCAGGAG ATCCCTTCAA GAGGCTGCAC CTTAATTTTT 8219 CTTGTCTGTG TGCAGGTCTT CATATAAACT TTACCAGGAA GAAGGGTGTG AGTTTGTTGT 8279 TTTTCTGTGT ATGGGCCTGG TCAGTGTAAA GTTTTATCCT TGATAGTCTA GTTACTATGA 8339 CCCTCCCCAC TTTTTTAAAA CCAGAAAAAG GTTTGGAATG TTGGAATGAC CAAGAGACAA 8399 GTTAACTCGT GCAAGAGCCA GTTACCCACC CACAGGTCCC CCTACTTCCT GCCAAGCATT 8459 CCATTGACTG CCTGTATGGA ACACATTTGT CCCAGATCTG AGCATTCTAG GCCTGTTTCA 8519 CTCACTCACC CAGCATATGA AACTAGTCTT AACTGTTGAG CCTTTCCTTT CATATCCACA 8579 GAAGACACTG TCTCAAATGT TGTACCCTTG CCATTTAGGA CTGAACTTTC CTTAGCCCAA 8639 GGGACCCAGT GACAGTTGTC TTCCGTTTGT CAGATGATCA GTCTCTACTG ATTATCTTGC 8699 TGCTTAAAGG CCTGCTCACC AATCTTTCTT TCACACCGTG TGGTCCGTGT TACTGGTATA 8759 CCCAGTATGT TCTCACTGAA GACATGGACT TTATATGTTC AAGTGCAGGA ATTGGAAAGT 8819 TGGACTTGTT TTCTATGATC CAAAACAGCC CTATAAGAAG GTTGGAAAAG GAGGAACTAT 8879 ATAGCAGCCT TTGCTATTTT CTGCTACCAT TTCTTTTCCT CTGAAGCGGC CATGACATTC 8939 CCTTTGGCAA CTAACGTAGA AACTCAACAG AACATTTTCC TTTCCTAGAG TCACCTTTTA 8999 GATGATAATG GACAACTATA GACTTGCTCA TTGTTCAGAC TGATTGCCCC TCACCTGAAT 9059 CCACTCTCTG TATTCATGCT CTTGGCAATT TCTTTGACTT TCTTTTAAGG GCAGAAGCAT 9119 TTTAGTTAAT TGTAGATAAA GAATAGTTTT CTTCCTCTTC TCCTTGGGCC AGTTAATAAT 9179 TGGTCCATGG CTACACTGCA ACTTCCGTCC AGTGCTGTGA TGCCCATGAC ACCTGCAAAA 9239 TAAGTTCTGC CTGGGCATTT TGTAGATATT AACAGGTGAA TTCCCGACTC TTTTGGTTTG 9299 AATGACAGTT CTCATTCCTT CTATGGCTGC AAGTATGCAT CAGTGCTTCC CACTTACCTG 9359 ATTTGTCTGT CGGTGGCCCC ATATGGAAAC CCTGCGTGTC TGTTGGCATA ATAGTTTACA 9419 AATGGTTTTT TCAGTCCTAT CCAAATTTAT TGAACCAACA AAAATAATTA CTTCTGCCCT 9479 GAGATAAGCA GATTAAGTTT GTTCATTCTC TGCTTTATTC TCTCCATGTG GCAACATTCT 9539 GTCAGCCTCT TTCATAGTGT GCAAACATTT TATCATTCTA AATGGTGACT CTCTGCCCTT 9599 GGACCCATTT ATTATTCACA GATGGGGAGA ACCTATCTGC ATGGACCCTC ACCATCCTCT 9659 GTGCAGCACA CACAGTGCAG GGAGCCAGTG GCGATGGCGA TGACTTTCTT CCCCTGGGAA 9719 TTCC 9723

Claims: - 146 - WHAT IS CLAIMED IS: 1. A pharmaceutical composition comprising a therapeutically effective amount of a Notch protein; and a pharmaceutically acceptable carrier. 2. The composition of claim 1 in which the Notch protein is a human Notch protein. 3. A pharmaceutical composition comprising a therapeutically effective amount of a protein, said protein comprising an amino acid sequence encoded by the DNA sequence depicted in Figure 8A (SEQ ID NO:5), 8B (SEQ ID NO:6), 8C (SEQ ID NO:7), 9A (SEQ ID NO: 8), or 9B (SEQ ID NO:9). which is able to be bound by an antibody to a Notch protein; and a pharmaceutically acceptable carrier. 4. A pharmaceutical composition comprising a therapeutically effective amount of a protein, said protein comprising a Notch amino acid sequence depicted in Figure 8A (SEQ ID NO:5), 8B (SEQ ID NO:6), 8C (SEQ ID NO:7), 9A (SEQ ID NO:8), or 9B (SEQ ID NO:9), which displays one or more functional activities associated with a full-length Notch protein; and a pharmaceutically acceptable carrier. 5. A pharmaceutical composition comprising a therapeutically effective amount of a protein, said protein comprising a fragment of a human Notch protein consisting essentially of the extracellular domain of the protein; and a pharmaceutically acceptable carrier. 6. A pharmaceutical composition comprising a therapeutically effective amount of a protein, said protein comprising a region of a Notch protein containing the EGF homologous repeats of the protein; and a pharmaceutically acceptable carrier. - 147 - 7. A pharmaceutical composition comprising a therapeutically effective amount of a fragment of a Notch protein lacking a portion of the EGF- homologous repeats of the protein, which fragment is able to be bound by an antibody to a Notch protein; and a pharmaceutically acceptable carrier. 8. A pharmaceutical composition comprising a therapeutically effective amount of a protein, said protein comprising a functionally active portion of a Notch protein; and a pharmaceutically acceptable carrier. 9. The composition of claim 8 in which the Notch protein is a human Notch protein. 10. A pharmaceutical composition comprising a therapeutically effective amount of a chimeric protein, said chimeric protein comprising a functionally active portion of a human Notch protein joined via a peptide bond to a sequence of a protein different from the Notch protein; and a pharmaceutically acceptable carrier. 11. The composition of claim 10 in which the functionally active portion of the Notch protein is encoded by the human cDNA sequence contained in plasmid hN3k as deposited with the ATCC and assigned accession number 68609, or encoded by the human cDNA sequence contained in plasmid hN5k as deposited with the ATCC and assigned accession number 68611. 12. A pharmaceutical composition comprising a therapeutically effective amount of a protein, said protein comprising the amino acid sequence depicted in Figure 10 (SEQ ID NO: 11); and a pharmaceutically acceptable carrier. 13. A pharmaceutical composition comprising a therapeutically effective amount of a protein, said protein comprising the amino acid sequence - 148 - depicted in Figure 11 (SEQ ID NO: 13); and a pharmaceutically acceptable carrier. 14. A pharmaceutical composition comprising a therapeutically effective amount of a protein, said protein comprising the portion of a human Notch protein with the greatest homology to the epidermal growth factor-like repeats 11 and 12 of the Drosophila Notch sequence as shown in Figure 4 (SEQ ID NO: 14); and a pharmaceutically acceptable carrier. 15. A pharmaceutical composition comprising a therapeutically effective amount of a derivative or analog of a Notch protein, which derivative or analog is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Delta protein expressed on the surface of a second cell; and a pharmaceutically acceptable carrier. 16. A pharmaceutical composition comprising a therapeutically effective amount of a chimeric protein, said chimeric protein comprising a Notch protein joined via a peptide bond to a protein sequence of a protein different from the Notch protein; and a pharmaceutically acceptable carrier. 17. A pharmaceutical composition comprising a therapeutically effective amount of a fragment of a Notch protein, which fragment is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Delta protein expressed on the surface of a second cell; and a pharmaceutically acceptable carrier. 18. A pharmaceutical composition comprising a therapeutically effective amount of a chimeric protein, said chimeric protein comprising a fragment of a Notch protein joined via a peptide bond to a protein sequence of a protein different from the Notch protein, which fragment is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Delta - 149 - protein expressed on the surface of a second cell; and a pharmaceutically acceptable carrier. 19. A pharmaceutical composition comprising a therapeutically effective amount of a protein, said protein comprising a derivative or analog of a Delta protein, which derivative or analog is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Notch protein expressed on the surface of a second cell; and a pharmaceutically acceptable carrier. 20. A pharmaceutical composition comprising a therapeutically effective amount of a chimeric protein, said chimeric protein comprising a fragment of a Delta protein joined via a peptide bond to a protein sequence of a protein different from the Delta protein, which fragment is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Notch protein expressed on the surface of a second cell; and a pharmaceutically acceptable carrier. 21. A pharmaceutical composition comprising a therapeutically effective amount of a protein, said protein comprising a derivative or analog of a Serrate protein, which derivative or analog is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Notch protein expressed on the surface of a second cell; and a pharmaceutically acceptable carrier. 22. A pharmaceutical composition comprising a therapeutically effective amount of a derivative or analog of a Notch protein, which derivative or analog is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a second protein expressed on the surface of a second cell, which second protein is selected from the group consisting of a Delta protein and a Serrate protein; and a pharmaceutically acceptable carrier. - 150 - 23. A pharmaceutical composition comprising a therapeutically effective amount of a nucleic acid encoding a Notch protein; and a pharmaceutically acceptable carrier. 24. A pharmaceutical composition comprising a therapeutically effective amount of a nucleic acid encoding a functionally active portion of a human Notch protein; and a pharmaceutically acceptable carrier. 25. A pharmaceutical composition comprising a therapeutically effective amount of a nucleic acid encoding the amino acid sequence depicted in Figure 10 (SEQ ID NO: 11); and a pharmaceutically acceptable carrier. 26. A pharmaceutical composition comprising a therapeutically effective amount of a nucleic acid encoding the amino acid sequence depicted in Figure 11 (SEQ ID NO: 13); and a pharmaceutically acceptable carrier. 27. A pharmaceutical composition comprising a therapeutically effective amount of a nucleic acid encoding a fragment of a Notch protein, which fragment is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Delta protein expressed on the surface of a second cell; and a pharmaceutically acceptable carrier. 28. A pharmaceutical composition comprising a therapeutically effective amount of a nucleic acid encoding a chimeric protein, said chimeric protein comprising a functionally active fragment of a human Notch protein joined via a peptide bond to a protein sequence of a protein different from the Notch protein; and a pharmaceutically acceptable carrier. 29. A pharmaceutical composition comprising a therapeutically effective amount of a nucleic acid encoding a fragment of a Delta protein, which fragment is characterized by the ability in vitro, when expressed on the surface of - 151 - a first cell, to bind to a Notch protein expressed on the surface of a second cell; and a pharmaceutically acceptable carrier. 30. A pharmaceutical composition comprising a therapeutically effective amount of a nucleic acid encoding a fragment of a Serrate protein, which fragment is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Notch protein expressed on the surface of a second cell; and a pharmaceutically acceptable carrier. 31. The composition of claim 24 in which the nucleic acid is a nucleic acid vector. 32. A pharmaceutical composition comprising a therapeutically effective amount of an antibody which binds to a Notch protein; and a pharmaceutically acceptable carrier. 33. A pharmaceutical composition comprising a therapeutically effective amount of a fragment or derivative of an antibody to a Notch protein containing the idiotype of the antibody; and a pharmaceutically acceptable carrier. 34. A method of treating or preventing a disease or disorder in a subject comprising administering to a subject in need of such treatment or prevention a therapeutically effective amount of a molecule which antagonizes the function of a Notch protein. 35. The method according to claim 34 in which the disease or disorder is a malignancy characterized by increased Notch activity or increased expression of a Notch protein or of a Notch derivative capable of being bound by an anti-Notch antibody, relative to said Notch activity or expression in an analogous non-malignant sample. - 152 - 36. The method according to claim 34 in which the disease or disorder is cervical cancer. 37. The method according to claim 34 in which the disease or disorder is breast cancer. 38. The method according to claim 34 in which the disease or disorder is colon cancer. 39. The method according to claim 35 in which the malignancy is selected from the group consisting of melanoma, seminoma, and lung cancer. 40. The method according to claim 35 in which the subject is a human. 41. The method according to claim 36, 37 or 38 in which the molecule is an antibody to Notch or a portion of said antibody containing the binding domain thereof. 42. The method according to claim 36, 37 or 38 in which the molecule is a protein consisting of at least the extracellular domain of a Notch protein or a portion thereof capable of binding to a Notch ligand. 43. The method according to claim 36, 37 or 38 in which the molecule is a protein consisting of at least the EGF homologous repeats of a Notch protein. 44. The method according to claim 36, 37 or 38 in which the molecule is a protein consisting of at least an adhesive fragment of a Notch protein. - 153 - 45. The method according to claim 36, 37 or 38 in which the molecule is an oligonucleotide which (a) consists of at least six nucleotides; (b) comprises a sequence complementary to at least a portion of an RNA transcript of a Notch gene; and (c) is hybridizable to the RNA transcript. 46. A method of treating or preventing a disease or disorder in a subject in need of such treatment or prevention comprising administering to the subject a therapeutically effective amount of a molecule which promotes the function of a Notch protein. 47. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a Notch protein. 48. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a functionally active portion of a Notch protein. 49. The method according to claim 47 in which the Notch protein is a human Notch protein. 50. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a chimeric protein, said protein comprising a functionally active portion of a Notch protein joined via a peptide bond to a protein sequence of a protein different from the Notch protein. 51. The method according to claim 49 in which the human Notch protein comprises the amino acid sequence depicted in Figure 10 (SEQ ID NO: 11) or Figure 11 (SEQ ID NO: 13). - 154 - 52. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a derivative or analog of a Notch protein, which derivative or analog is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a second protein expressed on the surface of a second cell, which second protein is selected from the group consisting of a Delta protein and a Serrate protein. 53. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a derivative or analog of a Delta protein, which derivative or analog is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Notch protein expressed on the surface of a second cell. 54. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a derivative or analog of a Serrate protein, which derivative or analog is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Notch protein expressed on the surface of a second cell. 55. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a nucleic acid encoding a Notch protein. 56. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a nucleic acid encoding a functionally active portion of a Notch protein. 57. The method according to claim 55 in which the subject is human and the Notch protein is a human Notch protein. - 155 - 58. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a nucleic acid encoding a fragment of a Notch protein, which fragment is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a second protein expressed on the surface of a second cell, which second protein is selected from the group consisting of a Delta protein and a Serrate protein. 59. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a nucleic acid encoding a fragment of a Delta protein, which fragment is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Notch protein expressed on the surface of a second cell. 60. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a nucleic acid encoding a fragment of a Serrate protein, which fragment is characterized by the ability in vitro, when expressed on the surface of a first cell, to bind to a Notch protein expressed on the surface of a second cell. 61. A method of treating or preventing a malignancy in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of antibody to a Notch protein. 62. The method according to claim 58 in which the antibody is monoclonal. 63. A method for treating a patient with a tumor, of a tumor type characterized by expression of a Notch gene, comprising administering to the patient an effective amount of an oligonucleotide, which oligonucleotide (a) consists of at least six nucleotides; (b) comprises a sequence complementary to at - 156 - least a portion of an RNA transcript of the Notch gene; and (c) is hybridizable to the RNA transcript. 64. The method according to claim 60 in which the patient is a human, and the Notch gene is a human gene. 65. An isolated oligonucleotide consisting of at least six nucleotides, and comprising a sequence complementary to at least a portion of an RNA transcript of a Notch gene, which oligonucleotide is hybridizable to the RNA transcript. 66. A pharmaceutical composition comprising the oligonucleotide of claim 65; and a pharmaceutically acceptable carrier. 67. A method of inhibiting the expression of a nucleic acid sequence encoding a Notch protein in a cell comprising providing the cell with an effective amount of the oligonucleotide of claim 65. 68. A method of diagnosing a disease or disorder characterized by an aberrant level of Notch protein or activity in a patient, comprising measuring the level of Notch protein expression or activity in a sample derived from the patient, in which an increase or decrease in Notch protein or activity in the patient sample relative to the level found in such a sample from a normal individual indicates the presence of the disease or disorder in the patient. 69. A method of diagnosing a malignancy characterized by an increased amou,_: of a Notch protein or of a Notch derivative capable of being bound by an anti-Notch antibody, comprising measuring the amount of a Notch protein or of a Notch derivative capable of being bound by an anti-Notch antibody, in a sample containing or suspected of containing malignant cells from a patient, in which an increase in the amount of the Notch protein or of the Notch - 157 - derivative capable of being bound by an anti-Notch antibody, in the sample, relative to said amount found in an analogous sample of non-malignant cells indicates the presence of the disease or disorder in the patient. 70. The method according to claim 69 in which the malignancy is cervical cancer. 71. The method according to claim 69 in which the malignancy is breast cancer. 72. The method according to claim 69 in which the malignancy is colon cancer. 73. The method according to claim 69 in which the malignancy is selected from the group consisting of melanoma, seminoma, and lung cancer. 74. The method according to claim 69 in which the amount of the Notch protein or derivative is measured by a method comprising contacting the sample with an anti-Notch antibody such that immunospecific binding can occur, and measuring the amount of any immunospecific binding of the antibody that occurs. 75. A method of treating or preventing a nervous system disorder in a subject comprising administering to a subject in need of such treatment or prevention an effective amount of a functionally active portion of a Notch protein. 76. A method of promoting tissue regeneration or repair in a subject comprising administering to a subject an effective amount of a functionally active portion of a Notch protein. - 158 - 77. A method of treating a benign dysproliferative disorder in a subject comprising administering to a subject in need of such treatment an effective amount of a functionally active portion of a Notch protein, in which the disorder is selected from the group consisting of cirrhosis of the liver, psoriasis, keloids, and baldness. 78. A substantially purified human Notch protein comprising the amino acid sequence encoded by the hN homolog as depicted in Figure 13 from amino acid numbers 1 through 2169 (SEQ ID NO: 19). 79. A substantially purified human Notch protein comprising the amino acid sequence encoded by the hN homolog as depicted in Figure 13 from amino acid numbers about 26 through 2169 (as contained in SEQ ID NO: 19). 80. A substantially purified protein comprising the extracellular domain of the mature human Notch protein encoded by the hN homolog, as depicted in Figure 13 from amino acid numbers about 26 through 1677 (as contained in SEQ ID NO: 19). 81. A substantially purified protein comprising the EGF homologous repeats of the mature human Notch protein encoded by the hN homolog, as depicted in Figure 13 from amino acid numbers 26 through 1413 (as contained in SEQ ID NO: 19). 82. A substantially purified protein comprising the EGF like repeats 11 and 12 of the mature human Notch protein encoded by the hN homolog, as depicted in Figure 13 (as contained in SEQ ID NO: 19). 83. A substantially purified protein consisting essentially of the extracellular domain of the mature human Notch protein encoded by the hN - 159 - homolog, as depicted in Figure 13 from amino acid numbers about 26 through 1677 (as contained in SEQ ID NO: 19). 84. A substantially purified nucleic acid encoding the protein of claim 78. 85. A substantially purified nucleic acid encoding the protein of claim 79. 86. A substantially purified nucleic acid encoding the protein of claim 80. 87. A substantially purified nucleic acid encoding the protein of claim 82. 88. The nucleic acid of claim 85 which is a DNA molecule comprising the sequence depicted in Figure 17 from nucleotide numbers 82 through 7419 (as contained in SEQ ID NO:21). 89. The nucleic acid of claim 80 in which the sequence encoding the extracellular domain is as presented in Figure 17 (as contained in SEQ ID NO:21). 90. A recombinant cell containing the nucleic acid of claim 84, 87 or 88. 91. The composition of claim 2 in which the Notch protein comprises the amino acid sequence encoded by the hN homolog as depicted in Figure 13 from amino acid numbers 26 through 2169 (as contained in SEQ ID NO: 19). - 160 - 92. A composition comprising a therapeutically effective amount of a Notch protein or Notch derivative, said derivative being capable of being bound by an anti-Notch antibody, for use as a medicament. 93. A composition comprising a therapeutically effective amount of a molecule which antagonizes the function of a Notch protein, for use as a medicament. 94. Use of a composition comprising a molecule which antagonizes the function of a Notch protein, for the manufacture of a medicament for the treatment of cervical cancer, breast cancer, or colon cancer.

Inventors: ARTAVANIS-TSAKONAS SPYRIDON, BLAUMUELLER CHRISTINE MARIE, FEHON RICHARD GRANT, ZAGOURAS PANAYIOTIS, ARTAVANIS-TSAKONAS, SPYRIDON, BLAUMUELLER, CHRISTINE, MARIE, FEHON, RICHARD, GRANT, ZAGOURAS, PANAYIOTIS
Applicants: ARTAVANIS TSAKONAS SPYRIDON, BLAUMUELLER CHRISTINE MARIE, FEHON RICHARD GRANT, UNIV YALE, ZAGOURAS PANAYIOTIS, YALE UNIVERSITY
Application year: 1993
Main classification: A61K31
IPC codes: A61K31, A61K38, A61K39, A61K48, A61P1, A61P17, A61P25, A61P35, A61P43, C07H21, C07K14, C12N1, C12N15, C12P21, C12R1, G01N33