HUNTINGTIN-ASSOCIATED PROTEIN

Abstract: Proteins which specifically bind to huntingtin, the product of the Huntington's Disease locus (HD), are used in assays for screening drug candidates. These proteins, termed huntingtin associated proteins, bind to huntingtins of both normal and Huntington's Disease patients. However, the binding is stronger to the huntingtins from the patients. The strength of binding correlates with the number of glutamine repeats in the huntingtin, which itself correlates with the time of disease onset.

Description: HUNTINGTIN-ASSOCIATED PROTEIN The U.S. Government has a paid-up license in this invention and the right in limited circumstances to require the patent owner to license others on reasonable terms as provided for by the terms of grants awarded by the National Institutes of Health. BACKGROUND OF THE INVENTION Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by an expanding glutamine repeat in a gene termed IT15 or huntingtii. 1 . Although this gene is widely expressed 2"9 and is required for normal development 10"12 , the pathology of HD is restricted to the brain, for reasons that remain poorly understood. The huntingtin gene product is expressed at similar levels in patients and controls, and the genetics of the disorder 13 14 suggest that the expansion of the polyglutamine repeat induces a toxic gain of function, perhaps through interactions with other cellular proteins 15"18 . There is a need in the art for identification of the cellular components which are involved with huntingtin so that the disease can be better understood, diagnosed, and treated. SUMMARY OF THE INVENTION It is an object of the invention to provide a method for identifying compounds which interfere with the binding of huntingtin associated protein to huntingtin. It is another object of the invention to provide methods for identifying compounds as candidate therapeutic agents. It is an object of the invention to provide a method for identifying compounds which diminish the binding of huntingtin associated protein to huntingtin. It is still another object of the invention to provide cells which can be used to identify compounds which interfere with the binding of huntingtin associated protein to huntingtin. It is yet another object of the invention to provide a method for determining the quantity of human huntingtin which binds to human huntingtin associated protein, or of human huntingtin associated protein which binds to human huntingtin. It is also an object of the invention to provide a method for screening for candidate drugs for treating, delaying onset, or preventing Huntington's Disease. It is an object of the invention to provide a yeast cell useful in the screening of candidate drugs. It is another object of the invention to provide cDNA segments encoding all or part of mammalian huntingtin associated proteins. It is yet another object of the invention to provide fusion proteins comprising huntingtin associated proteins. It is still another object of the invention to provide isolated mammalian proteins which are huntingtin associated proteins. These and other objects of the invention are provided by one or more embodiments as described below. In one embodiment, a method for identifying compounds which interfere with the binding of huntingtin associated protein to huntingtin is provided. The compounds identified are candidate therapeutic agents. The method comprises the steps of: contacting: a first protein; a second protein; and a compound to be tested for its capacity to interfere with binding of said first and second proteins to each other; wherein the first protein comprises huntingtin associated protein and the second protein comprises huntingtin or the first protein comprises huntingtin and the second protein comprises huntingtin associated protein; and determining the quantity of the first protein which is bound to, is displaced from, or is prevented from binding to, the second protein, wherein a compound which diminishes the quantity of the first protein bound to the second protein, or which displaces first protein bound to the second protein, or which prevents first protein from binding to the second protein, is identified as a candidate therapeutic agent. In another embodiment, a second method for identifying compounds which interfere with the binding of huntingtin associated protein to huntingtin is provided. The identified compounds are candidate therapeutic agents. The method comprises the steps of: contacting: a first polypeptide; a second polypeptide; and a compound to be tested for its capacity to interfere with binding of said first and second polypeptides to each other; wherein at least one of said first and said second polypeptides is a polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin or huntingtin associated protein proteins, wherein each of said polypeptides contain a sufficient portion of huntingtin or huntingtin associated protein proteins to bind to the other polypeptide; wherein when said first polypeptide is huntingtin or a polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin, then said second polypeptide is huntingtin associated protein or a polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin associated protein; and when said first polypeptide is huntingtin associated protein or a polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin associated protein, then said second polypeptide is huntingtin or a polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin; and determining the quantity of the first polypeptide which is bound to, is displaced from, or is prevented from binding to, the second polypeptide, wherein a compound which diminishes the quantity of the first protein, or which prevents first protein from binding to the second protein, is identified as a candidate therapeutic agent. In yet another embodiment of the invention a third method of identifying compounds which interfere with the binding of human huntingtin associated protein to human huntingtin is provided. The method comprises: providing a cell which comprises three recombinant DNA constructs, said first construct encoding a first polypeptide fused to a sequence- specific DNA-binding domain, said second construct encoding a second polypeptide fused to a transcriptional activation domain, said third construct comprising a reporter gene downstream from a DNA element which is recognized by said sequence-specific DNA-binding domain, wherein said first polypeptide is a huntingtin associated protein polypeptide and said second protein is a huntingtin polypeptide, or said first polypeptide is a huntingtin polypeptide and said second polypeptide is a huntingtin associated protein polypeptide; wherein said polypeptides contain a sufficient portion of huntingtin and huntingtin associated protein proteins to bind to the other polypeptide; contacting said cell with a compound to be tested for its capacity to inhibit binding of huntingtin to huntingtin associated protein; determining the amount of expression of the reporter gene in the presence of said compound. According to another embodiment a cell is provided for screening compounds. The cell comprises three recombinant DNA constructs, said first construct encoding a first polypeptide fused to a sequence-specific DNA-binding domain, said second construct encoding a second polypeptide fused to a transcriptional activation domain, said third construct comprising a reporter gene downstream from a DNA element which is recognized by said sequence-specific DNA-binding domain, wherein said first polypeptide is an huntingtin associated protein polypeptide and said second protein is a huntingtin polypeptide, or said first polypeptide is a huntingtin polypeptide and said second polypeptide is an huntingtin associated protein polypeptide; wherein said polypeptides contain a sufficient portion of huntingtin and huntingtin associated protein proteins to bind to the other polypeptide. According to another embodiment a method is provided for determining the quantity of human huntingtin which binds to human huntingtin associated protein, or of human huntingtin associated protein which binds to human huntingtin. The method comprises: contacting: a first protein and a second protein, wherein the first protein is human huntingtin associated protein and the second protein is human huntingtin or the first protein is human huntingtin and the second protein is human huntingtin associated protein; and determining the quantity of the first protein which is bound to the second protein. In yet another embodiment of the invention, a method of screening for candidate drugs for treating, delaying onset, or preventing Huntington's Disease is provided. The method comprises the steps of: contacting a compound to be tested with a yeast cell comprising: two fused gene constructs wherein a first construct comprises a yeast GAL-4 binding domain and a domain selected from the group consisting of: a glutamine repeat-containing domain of a huntingtin gene and a huntingtin binding domain of a huntingtin associated protein gene, and wherein a second construct comprises a yeast GAL-4 activation domain, and a domain selected from the group consisting of: a glutamine repeat-containing domain of a huntingtin gene and a huntingtin binding domain of a huntingtin associated protein gene, wherein when the first construct comprises a glutamine repeat-containing domain of a huntingtin gene, the second construct comprises a huntingtin binding domain of a huntingtin associated protein gene, and when the second construct comprises a glutamine repeat-containing domain of a huntingtin gene, the first construct comprises a huntingtin binding domain of a huntingtin associated protein gene; and a /3-galactosidase reporter gene under the control of a yeast GAL-4 promoter, which is activated by the gene products of said two fused gene constructs, and selecting those compounds which diminish the expression of β- galactosidase by said cells. In another embodiment of the invention a yeast cell is provided. The yeast cell comprises: two fused gene constructs and a 0-galactosidase reporter gene under the control of a yeast GAL-4 promoter, wherein a first construct comprises a yeast GAL-4 binding domain and a domain selected from the group consisting of: a glutamine repeat- containing domain of a huntingtin gene and a huntingtin binding domain of a huntingtin associated protein gene, and a second construct comprises a yeast GAL-4 activation domain, and a domain selected from the group consisting of: a glutamine repeat-containing domain of a huntingtin gene and a huntingtin binding domain of a huntingtin associated protein gene, wherein when the first construct comprises a glutamine repeat-containing domain of a huntingtin gene, the second construct comprises a huntingtin binding domain of a huntingtin associated protein gene, and when the second construct comprises a glutamine repeat- containing domain of a huntingtin gene, the first construct comprises a huntingtin binding domain of a huntingtin associated protein gene; and said 3-galactosidase reporter gene is activated by the gene products of said two fused gene constructs. According to still another embodiment of the invention a cDNA segment is provided. The cDNA segment encodes at least 8, 12, or preferably 15 contiguous amino acids of a mammalian huntingtin associated protein. In yet another embodiment of the invention a cDNA segment is provided which consists of at least 20 or preferably 30 or 40 contiguous nucleotides of a mammalian gene encoding huntingtin associated protein. In another embodiment of the invention a fusion protein is provided. It consists of all or a portion of a first and a second protein, wherein said second protein is a mammalian huntingtin associated protein, and the portion of said mammalian huntingtin associated protein is sufficient to bind to huntingtin. In still another embodiment of the invention an isolated mammalian protein is provided. It is a huntington associated protein. These and other embodiments of the invention provide the art with techniques and tools for identifying drugs for the amelioration of Huntington's Disease. BRIEF DESCRIPTION OF THE DRAWINGS Figures la-lf: Association of rat HAP1 with huntingtin. (Fig. la) Filter assays for 0-galactosidase activity in transformed yeast with proteins fused to the GAL-4 binding domain (BD) and the activation domain(AD) (BD construct + AD construct). HAP1: AD construct of rHAPl. Only co-expression of rHAPl plus the N-terminal portions of huntingtin containing either 23 or 44 glutamine repeats (HD23Q or HD44Q) activate the /3-galactosidase reporter gene (blue). Co-transformation with c-Fos and c-Jun (Fos+Jun) served as the positive control. (Rg. lb) Liquid assays of interactions of rHAPl with various proteins in yeast cells. Atrophin (21Q) is the GAL-4 binding domain construct containing 263 amino acids of atrophin- 1 with 21 glutamine repeats. (Rg. lc) Coomassie blue staining of GST-ENAC, GST and GST-HAP1 (rHAPl) fusion proteins. ENAC is the N-terminal portion (107 amino acids) of a subunit of the amiloride-sensitive sodium channel expressed predominately in colon, kidney and tongue but not in brain 24,25 . (Rg. Id) Western blot analysis of huntingtins from 293 cells transfected with an HD cDNA using anti-peptide antibody for the huntingtin (AP-81) 4 . (Rg. le) Binding of GST-HAP1 to huntingtins in transfected 293 cells. (Rg. If) Inputs of huntingtins from lymphoblasts with 22/19; 44 /20; and 82/19 glutamine repeats, respectively, were reacted with the GST-HAP 1 fusion protein and Western blot (Fig. lg) reveals the amount of huntingtin bound to GST-HAP 1. Rgures 2a-2c. Schematic diagram and deduced amino acid sequences of HAP1 and HLP1. (Rg. 2a) The open reading frame is indicated as a box. A black box corresponds to the region of rHAPl cDNA isolated from the yeast two hybrid screening. rHAPl -A and rHAPl -B were isolated from rat brain cDNA libraries and have identical sequences in their overlapping regions, with different amino acids in the C-terminus (shown by shaded boxes at the C-terminus). An acidic amino acid rich region is indicated as a shaded box. rHAPl -A contains the entire translated region, while rHAPl-B contains the same sequence of the untranslated region on the end 3'. The region of partial cDNA of human HLP1 or HAP1 isolated by RT-PCR are also shown. (Fig. 2b) The first ATG in rHAPl -A is used as a putative start codon and deduced amino acid sequences of rHAPl -A and rHAPl-B are shown. The acidic amino acid rich region is underlined and the original sequence of rHAPl isolated from the yeast two hybrid system is in the bold type. (Rg. 2c) Alignment of partial human HLP1 and HAP1 amino acid sequences with rHAPl A. Amino acid sequences derived from PCR primers are underlined. Amino acid residues identical to rHAPl are indicated with vertical lines. Rgures 3a-3c. Expression of HAP1. (Rg. 3a) Northern blot analysis of rat HAP-1 mRNA (Fig. 3b) and RT-PCR analysis of expression of human HAP1 using primers spanning the binding region. St.N. : subthalamic nucleus, Cd.: Caudate, Ctx. Cerebral cortex, Hip.: Hippocampus, Cereb.: cerebellum, Genomic: human genomic DNA, dH2O.: no RNA input. hHAPl: hHAPl plasmid cDNA. (Rg. 3c) Immunoblot analysis of HAP1 protein. Human Ctx: Human cerebral Cortex. Rgures 4a-4b . Co-immunoprecipitation of (Fig.4a) transfected rH AP 1 -A and endogenous human huntingtins in HEK 293 cells and (Fig. 4b) endogenous rat HAP1 and huntingtins. 293 cells: untransfected cells; Lysates: cell extracts from transfected 293 cells with rHAPl-A cDNA; AP81 : with purified antibody (AP-81) for huntingtin; IP 3 R-1: with antibody for IP 3 receptor type 1; AP81+ peptide: with AP81 preabsorbed with the peptide immunogen. HAPl-Ab, rat HAP1 antibodies. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS It is a finding of the present inventors that huntingtin associated protein (HAP1) binds to the protein product of mammalian Huntington's Disease loci, termed herein, huntingtin. The binding is enhanced by an expanded polyglutamine repeat in huntingtin, the length of which is also known to correlate with the time of disease onset 1 * *21 . In addition, HAP1 protein, in contrast to huntingtin itself, is expressed selectively in the brain. The normal function of huntingtin is unknown, though it has been proposed to play a role in microtubule mediated transport or vesicle function 7,9 . The selective expression of HAP 1 in brain, in contrast to the ubiquitous distribution of HD gene expression, suggests that HAP1 may contribute to the brain-specific pathology of HD. The pathology of HD involves most or all of the brain in advanced cases, though some regions show striking early selective vulnerability. Though rat HAP1 (rHAPl) is highly expressed in many regions of rat brain, the message for human HAP1 (hHAPl) can be detected most consistently by RT-PCR in the human brain regions (caudate and cortex) that are most affected in HD. Thus, the levels of expression of human HAP1 in specific cell populations may influence their vulnerability. The screening methods of the present invention all rely on the principle of interference in the binding of huntingtin and huntingtin associated protein by a compound. Thus any assay format for measuring protein-protein interactions that is known in the art can profitably be used. For example, both in vitro and in vivo tests can be used. One of the proteins can be immobilized, for example, on a microtiter dish, or on a column packing matrix. Immobilization can be direct or indirect, e.g. , through a domain of a fused protein. Assays can employ radiolabels, enzyme assays, antibodies, florescent labels, etc. Binding interactions can be assessed by measuring the bound or the unbound fraction. According to some embodiments of the invention, the entire huntingtin or huntingtin associated protein is used, as is, or fused to another polypeptide domain or protein. Alternatively, polypeptide portions of either huntingtin or huntingtin associated protein can be used, so long as they contain the portions of huntingtin and huntingtin associated protein which are required for binding to each other. Suitable portions of human huntingtin associated protein for use include amino acids 105-136, 4-136, 50-136, and 75-136. Suitable portions of huntingtin for use include amino acids 1-230 and 1-930, although the minimal amount has not been determined. It is believed that the glutamine repeat units are necessary for binding, however, flanking amino acids may also be necessary for proper binding conformation and/or specificity. In vivo assays such as the GAL4 based assays described by Fields et al. 28 can be used. Such assays employ fusion proteins of the two interacting proteins of interest. One partner is fused to an activation domain and one partner is fused to a DNA binding domain. Neither domain by itself will activate transcription of a suitable reporter gene. However, when the two domains are brought into proximity, such as by the interaction of the two interacting proteins, then the reporter's transcription is activated. Other suitable systems have been developed. Spencer et al. Science, 262, 1019-1024 (1993) developed a system which relies on the interaction of FKBP12 and FK 1012. Fearon et al. , Proc. Natl. Acad. Sci. USA, 89, 7958-7962 (1992) teaches a system which is based on yeast GAL4 but which can be used in mammalian cells. Reporter genes which are used are preferably those whose expression can be quantitatively or semi-quantitatively assayed, including drug resistance enzymes and anabolic enzymes. Both the his3 and the 3-galactosidase genes can be used to advantage. In vivo assays may be preferable to in vitro assays because they require that the compound being tested penetrate the cells and locate the appropriate target proteins. However, both types of methods may be used, either individually or sequentially. Candidate drugs are identified as those which inhibit the binding of huntingtin to huntingtin associated protein. Such inhibitory molecules can be identified by screening for interference of the huntingtin/huntingtin associated protein interaction where one of the binding partners is bound to a solid support and the other partner is labeled. Antibodies specific for epitopes on huntingtin or huntingtin associated protein which are involved in the binding interaction will interfere with such binding. Solid supports which may be used include any polymers which are known to bind proteins. The support may be in the form of a filter, column packing matrix, beads, microtiter dish, and the like. Labeling of proteins can be accomplished according to any technique known in the art. Radiolabels, enzymatic labels, and fluorescent labels can be used advantageously. Alternatively, both huntingtin and huntingtin associated protein may be in solution and bound molecules separated from unbound subsequently. Any separation technique known in the art may be employed, including immunoprecipitation or immunoaffinity separation with an antibody specific for the unlabeled binding partner. It has been found that amino acid residues 1-230 of huntingtin (SEQ ID NO: 8) are sufficient for the interaction of huntingtin and huntingtin associated protein. However, some of these residues may not be necessary for binding. In addition, it has been found that residues 1-136 of human huntingtin associated protein are sufficient for binding to huntingtin. Again, less may be required. Loss of residues corresponding to residues 105-136 leads to non-binding proteins, as demonstrated by hHLPl. Thus it is presumed that some of the amino acids in the 105-136 region are necessary. Additional residues in the 1-105 region may also be required. According to another aspect of the invention, the binding of huntingtin and huntingtin associated protein can be quantitatively evaluated in the absence of test compounds. Such assays can be used to determine if a biological sample contains a mutant huntingtin protein. Such assays can also be used to determine if a particular tissue expresses huntingtin associated proteins. cDNA molecules and segments are also provided by the present invention. A segment may, for example, be covalently joined to a vector or to another gene. The cDNA molecules and segments of the present invention are those which encode huntingtin associated proteins. Preferably they encode at least 12 contiguous amino acids of a mammalian huntingtin associated protein. In some instances, such as in the preparation of gene primers and probes, the use of a segment consisting of as little as 17, 20, 25, or 30 nucleotides may be sufficient to obtain specific binding. Such segments may also be sufficient to encode epitopes useful in the generation of antibodies which specifically bind to huntingtin associated proteins. Immunization of experimental animals to generate polyclonal antisera is well-known in the art. Manipulation of such immunized animals to obtain monoclonal antisera is also well-known and contemplated by the invention. Huntingtin associated proteins are also contemplated by the present invention. Such proteins may be the full-length protein, as found in vivo, or may be portions of the protein which can be used as polypeptide immunogens, or fused to other protein domains. The full-length proteins of the present invention are isolated from other cellular components, such as DNA, RNA, lipids, and often other proteins. Fusion proteins can be made, as are known in the art, using recombinant DNA techniques and expression of fused genes in cells. Examples Example 1 This example demonstrates the isolation of the gene encoding huntingtin associated protein from rat. A cDNA encoding the first 230 amino acids of huntingtin containing 44 glutamine repeats was ligated to the GAL-4 binding domain in plasmid vector pPC97 for yeast two hybrid screening with a rat brain cDNA library. This region was chosen because it includes the portion of the protein in which a glutamine repeat is expanded in HD 1 . Overall, longer expansions are associated with earlier age of onset. Alleles with 44 repeats are among the most common in the expanded range, and can cause onset of the disorder at ages ranging from the 20s to the 60s 19"21 . Screening of approximately 100,000 colonies resulted in one positive clone. This clone (rHAPl -BR, rat huntingtin Associated Protein 1 Binding Region) had an insert of 507 base pairs with no homology to known genes. Co-transformation of yeast with rHAPl plus the HD constructs resulted in blue colonies which were not evident following co-transformation with either vector alone or with unrelated fusion proteins in either vector (Fig. la). The interaction was positive using the HD fusion protein containing either a normal repeat length (23 glutamines) or an expanded repeat (44 glutamines). The interaction of rHAPl with huntingtin was stronger with the 44 glutamine repeats than with the 23 repeats using a semi-quantitative liquid assay (Fig. lb). To determine whether the interaction is specific to huntingtin or would be positive for another protein with a polyglutamine tract, we performed a similar experiment using rHAPl plus an atrophin- 1 construct, which contains a 21 glutamine repeat 22 . This showed no evidence for interaction in either filter or liquid assays. Methods'. The partial length of HD cDNAs were isolated from a human brain cortex cDNA library (Stratagene) using a CAG repeat oligonucleotide probe 16 or PCR with the first strand cDNA from a HD brain tissue. A cDNA construct containing the first 230 amino acids with 44 or 23 glutamine repeats was in-frame fused with the GAL4 DNA binding domain of plasmid PPC97 27 . An adult rat hippocampus cDNA library was constructed using plasmid pPC86 containing the GAL4 activation domain. Transformations of a yeast strain PCY2 and filter assays of 0-galactosidase activity were performed according to the methods described previously 27 and the controls were included as described by Fields 28 . The liquid assay to quantify β-galactosidase activity is as previously described 29 . Example 2 This example demonstrates an in vitro binding assay using huntingtin and huntingtin associated protein fusion proteins. It also demonstrates the correlation between strength of binding and number of glutamine repeats in the huntingtin protein. In addition, it demonstrates that rat huntingtin associated protein binds to human huntingtin. In vitro binding assays were then performed using glutathione-S- transferase (GST) fusion proteins (Fig. lc). An HD cDNA encoding the N-terminal region (930 amino acids with 44 repeats) was fused in frame to an expression vector pEBVHIS (Invitrogen). Transfection of this construct into HEK-293 cells resulted in a band (131 kDa) that was recognized by a huntingtin specific antibody 4 (Fig. Id). The endogenous protein (350 kDa) could be seen as well. When an extract from cells transfected with the N-terminal huntingtin construct was incubated with GST-HAP 1 protein linked to agarose beads, both the native huntingtin and the transfected huntingtin with the expanded repeat (44 glutamines) were specifically retained on the beads (Figs, lc, Id, le). A similar experiment was performed using lymphoblastoid cell lines from a normal individual and HD patients with varying lengths of glutamine repeats (Fig. If). Cell extracts containing the huntingtin with 82 repeats yielded the strongest binding compared to the huntingtin with 44 repeats, and both proteins showed increased binding to GST-HAP1 compared to the protein from the normal individual (19-22 repeats) (Fig. lg). Alleles of > 70 repeats are in the severely expanded range and nearly always cause HD with juvenile onset. Methods'. rHAPl cDNA (507 bp) isolated from the yeast two hybrid screening was inserted into pGEX-2T vector (Pharmacia) and GST fusion protein expression and purification were essentially as previously described 30 . The sources containing huntingtins were 293 cells transfected with the cDNA encoding the partial N-terminal portion (930 amino acids with 44 glutamine repeats) of huntingtin fused in frame to pEBVHIS vector (Invitrogen) and transformed lymphocytes (lymphoblast) from a normal individual and HD patients. The 293 cells from a 10 cm dish or 100 ml lymphoblast cultures were lysed in 1.5 ml binding buffer (1 % Triton, 160 mM NaCl, 50 mM Hepes (pH 7.4), 2.5 mM MgCl, 1.5 mM CaCl and 2.5 mM KCl, ImM PMSF, and 2 μg/ml each of pepstatin, leupeptin and aprotinin). Cell lysates were pre-incubated with agarose beads containing about 0.5-1 mg GST protein at 4°C for 30 min, clarified by centrifugation at 10,000xg for 10 min at 4°C, then incubated with agarose beads containing 0.5 μg GST-HAP 1 or other fusion proteins at 4°C for 1 hr. Agarose beads were washed three times with 1 ml binding buffer. Proteins eluted from beads with SDS-sample buffer were detected in Western blot analysis with huntingtin antibodies AP-81. Example 3 This example demonstrates the cloning of two full-length cDNAs encoding huntingtin associated protein. The cDNA insert from the rHAPl clone was used to screen rat brain cDNA libraries to obtain a full length cDNA sequence. rHAPl-A and rHAPl-B represent two groups of cDNAs that display different sequences in the C-terminal region (Figs. 2a, 2b). There are no significant homologies to known genes in the database. Both proteins are highly hydrophilic with no hydrophobic region. Their calculated PI value is about 4.67 with a high percentage of charged amino acids, including a glutamate and aspartate rich region (Asp 217 - Asp 237 ). Methods: The cDNA for rat HAP1 (507 bp) isolated from the yeast two hybrid screening was labeled with 32 P as a probe to screen rat brain cDNA libraries (Stratagene). rHAPl-A and rHAPl-B represent two groups of cDNA (from a total of 30) with different sequences at the C-terminus. Six cDNAs have been sequenced on both strands and amino acid sequences of rHAPl-A and rHAPl-B are shown. Example 4 This example demonstrates the isolation of genes related to huntingtin associated protein from human. The encoded proteins do not bind to huntingtin. We then used a PCR approach to identify hHAPl from human caudate tissue, with a variety of primers based on the sequences of both ends of putative binding region (rHAPl). Six types of subclones of PCR products contain 5' and 3' regions with about 70% amino acid identity to rHAPl, but display different deletions or insertions in the region corresponding to the midportion of rHAPl. These cDNAs are termed hHLP (human HAP-like protein) because they all lack the midportion of rHAPl (between 371 Gly and 421 Asp) (Fig. 2c) and were unable to generate blue colonies when introduced into yeast with HD cDNA. Methods: RT-PCR with human caudate RNA was performed to identify human HLP cDNA. hHLPl was isolated with the sense primer gatgatctccttcagctctac (DDLLQLY) and antisense primer ccagttggacagttgctggaggac (VLQQLS). cDNA products were then subcloned and sequenced. Example 5 This example demonstrates the isolation of a cDNA fragment from human caudate tissue which is highly homologous to rat huntingtin associated protein and which binds to huntingtin. We obtained a cDNA fragment from human caudate tissue, which contains 94% nucleic acid identity (375 bp) to rHAPl with 96% amino acid identity and is termed hHAPl (human HAP1, Fig. 2c). Co-transformation of yeast with hHAPl cDNA plus the HD constructs (44 or 23 repeats) produced blue colonies with similar intensity to those with rHAPl cDNA (data not shown), indicating that the midportion of HAP1 is crucial for interactions with the HD protein. Southern analysis using human genomic DNA is consistent with the existence of at least two separate genes related to rHAPl (data not shown). Methods: The sense oligonucleotide primer accactgcccacagctggaag and the antisense primer catagtgggtgacagaacct that correspond to rHAPl amino acid sequence HCPQLE and GSVTHY respectively, were used to amplify human caudate cDNA at 95 °C for 1 min, 55° C for 1 min then 72 °C for 1 min for 30 cycles . cDNA were amplified again using the sense primer agcagaagctgaagctgctgga and antisense primer agaacctgcaggcattgagg that encode amino acid sequences KLKLLE and SMPAGS respectively, under the same PCR condition. Example 6 This example demonstrates that mRNA encoding huntingtin associated protein is highly expressed in the brain, but not in several peripheral tissues. Northern blot analysis of the expression of the rHAPl (Fig. 3a) shows transcripts of approximately 4.0 kb in rat brain, with the suggestion of two partially resolved bands. Expression is high in many brain regions. By contrast there is no detectable signal in several peripheral tissues (Fig. 3a). To confirm the expression of hHAPl in human brain, we conducted RT-PCR analysis with primers derived from hHAPl (Fig. 3b). We obtained PCR products with the expected size (320 bp), that could be detected by hybridization with a hHAPl cDNA probe. This form is detected in subthalamic nucleus, caudate, cerebral cortex, and fetal brain. Methods: Twenty μg total RNA from each tissue and the 32 P labeled cDNA probe of rHAPl were used in Northern blot analysis and the blot was exposed to X-ray film for 24 hr. For the human RT-PCR analysis (Rg. 3b), PCR products obtained using the sense primer atgctcattctggagtgtgtg (MLILECV) and antisense primer agaacctgcaggcattgagg (SMPAGS) at an annealing temperature 62 °C for 35 cycles were separated on 1 % agarose gel, transferred onto a nitrocellulose membrane and hybridized with the 32 P labeled cDNA probe of hHAPl under a high stringency conditions. One of five independent RT-PCR assays with commercial human cDNA (Clontech) or our synthesized cDNA is shown in (Rg. 3b). This experiment is representative, but other experiments did yield a detectable band in other brain regions such as hippocampus (Hip). Example 7 This example demonstrates that huntingtin associated protein is expressed in the brain but not in peripheral tissues, confirming the findings with mRNA expression. Affinity purified antibodies raised against GST-HAP 1 were used to confirm expression of the rHAPl protein. A construct containing the full length protein of rHAPl-A or rHAPl-B was transfected into HEK-293 cells. Western blots of rat whole brain contain three immunoreactive bands (Fig. 3c). The middle band is the same molecular mass as the transfected rHAPl-A protein (75 kDa) in 293 cells. In vitro translation of rHAPl-A cDNA also yields a product of the same apparent molecular weight (75 kDa, data not shown). The upper band (85 kDa) corresponds to the transfected rHAPl-B. The lowest, 68-70 kDa, is the size predicted for rHAPl-A or rHAPl-B cDNA and is present in both rat and human brain. By contrast, several peripheral tissues from rat and human showed no immunoreactivity (Fig. 3c). The immunoreactivity of all three bands could be eliminated by preabsorption of antibodies with the antigen (data not shown). The three bands in the rat brain might reflect alternative splicing, which would fit with the existence of two distinct cDNA sequences for rHAPl. The variety of HAP cDNA homologues obtained by PCR suggests the existence of multiple human HAP1 or HAP-like proteins (HLP). Antibodies for rHAPl might be unable to distinguish them because of their similar mobilities in the gel or different expression levels. Post-translational modification may also play a role and fit with the molecular weights of the upper two bands being greater than the predicted molecular weights. Methods: For the Western blots (Fig. 3c), tissues were homogenized in PBS buffer with protease inhibitors as described in Fig. 2 and about 75 mg proteins were used in Western blot except for transfected cells (lμg). HEK-293 cells were transfected with a pCIS-2 expression vector (Genentech) containing a rat HAP1 cDNA construct using the Ca + + -phosphate precipitation method. The N-terminal sequence of rHAPl-A was inserted into rHAPl-B using the Neo I restriction site to generate the full length protein of rHAPl-B. The GST-HAP 1 fusion protein was used as an antigen for production of antiserum from rabbits (HRP Inc.). The antiserum (1.5 ml) was incubated with 20 mg GST-HAP 1 antigen transferred onto the nitrocellulose. The nitrocellulose was washed with PBS, and antibodies were eluted with 1 ml 100 mM glycine (pH 2.8). The elution fluid was dialyzed against PBS overnight. Purified antiserum (1:2000) or preabsorbed antiserum in PBS containing 3% BSA was used for Western blot analysis with an enhanced chemiluminescence kit (Amersham). Example 8 This example demonstrates that, like the portion of huntingtin associated protein, full-length huntingtin associated protein interacts with huntingtin in vivo. To confirm that the full length rHAPl interacts with huntingtin in vivo, we conducted co-immunoprecipitation experiments. rHAPl-A protein transfected in 293 cells could be precipitated specifically with affinity purified antibody to huntingtin, AP81, and this precipitation could be blocked by the peptide antigen (Fig 4a). Using a different peptide antibody for huntingtin (AP78), we obtained similar results (data not shown). Using whole rat brain, we also observed coprecipitation of both rat HAP1 and huntingtins with AP78 or AP81 (Fig. 4b). We have not yet clearly shown co-immunoprecipitation from human brain tissue, which may reflect the complex mixture of hHAPl and HLPs and their possibly different tissue localization. Another factor may be the difficulty of using human postmortem brain tissues. The yeast two hybrid assays and the in vitro binding data show that the association between HAP1 and huntingtin is enhanced by increasing lengths of the glutamine repeat. Binding may be influenced by the flanking amino acids, since there is no binding to the atrophin- 1 protein which contains essentially the same number of glutamines (21) as the HD construct (23 glutamines). The affinity of the interaction may be increased by an altered conformation of the huntingtin due to the expanded glutamine repeat. The enhanced association of HAP1 with a huntingtin containing an expanded polyglutamine repeat could contribute to the pathophysiology of HD, consistent with the toxic gain of function hypothesis. Methods: Transfected cells with rHAPl-A cDNA from a 10 cm plate or whole rat brain (0.15 g/wt) were lysed in 1.5 ml binding buffer as for GST fusion protein binding experiments, and 750 ml lysate was incubated with 100 ml antibodies linked beads (50% slurry) at 4 °C for 1 hr. Affinity purified antibodies (AP-78 or AP81, 20 mg) were coupled to 1 ml protein -A agarose beads using 100 mM dimethyl pimelimidate. AP81 linked beads pre-absorbed with peptide immunogen (20 μg/ml) overnight or antibodies linked beads for IP3 receptor type-3 were used as the controls. Proteins were eluted from beads with 100 mM glycine (pH 2.8) following washes with the binding buffer and eluted proteins were detected on Western blots with purified rHAPl antibodies (1:2000). References: 1. Huntington's Disease Collaborative Research Group. Cell 72, 971-983 (1993) 2. Li, S-H., et al. Neuron 11, 985-993 (1993) 3. Strong, T. V., et al. Nature Genet. 5, 259-265 (1993) 4. Sharp, A.H., et al. Neuron 14, 1065-1074 (1995) 5. Schilling, G., et al. Hum. Mol. Genet. 4, 1365-1371 (1995) 6. Trottier, Y., et al. Nature Genet. 10, 104-110 (1995) 7. DiFiglia, M., et al. Neuron 14 1075-1081 (1995) 8. Persichetti, F., et al. Mol. Med. 1, 374-383 (1995) 9. Gutekunst, C.-A., et al. Proc. Natl. Acad. Sci.. in press 10. Nasir, J., Florescoet al. Cell 81, 811-823 (1995). 11. Duyao, M.P., et al. Science 269, 407-410 (1995) 12. Zeitlin, S., Liu, J.-P., Chapman, D.L., Papaioannou, V.E., and Efstrtiatis, A. , Nature Genet. 10, in press 13. Wexler, N.S., et al. Nature 326, 194-197 (1987) 14. Ambrose, CM., et al. Somat. Cell Mol. Genet. 20, 27-38 (1995) 15. Gusella, J.F., MacDonald, M.E., Ambrose, CM. and Duyao, M.P. Arch. Neurol. 50, 1157-1163 (1993) 16. Ross, C.A., Mclnnis, M.G., Margolis, R.L., Antonarakis, S.E. and Li, S-H. Trends Neurosci . 16. 254-260 f!993. 17. Albin, R.L., and Tagle, D.A. Trends Neurosci. 18, 11-14 (1995) 18. Ross, CA. Neuron 15, 493-496, (1995) 19. Duyao, M, et al. Nature Genetics 4, 387-392 (1993) 20. Andrew, S.E., et al. Nature Genet. 4. 398-403 (1993. 21. Stine, O.C et al. Hum. Mol. Genet. 2, 1547-1549 (1993) 22. Margolis, R.L., Li, S-H. and Ross, CA. Mol. Brain Res, in press, (1995) 23. Beal, M.F. Ann. Neurol. 38, 357-366 (1995) 24. Canessa, C. M., Horisberger, J. D. and Rossier, B. C Nature 361, 467-470. (1993) 25. Li, X.J., Blackshaw, S. and Snyder, S.H. Proc. Natl. Acad. Sci. USA 91, 1814-1818 (1994) 26. Li, S-H., Mclnnis, M.G., Margolis, R.L., Antonarakis, S.E. and Ross, CA . Genomics 16, 572-579 (1993) 27. Chevray, P. M. and Nathans, D. Proc Natl. Acad. Sci. US A 89, 5789-5793 (1992) 28. Fields, S. and Sternglanz, R. Trends Genet 10, 286-292 (1994) 29. Guarente, L. Methods Enzvmol. 101, 181-91 (1983) 30. Guan, K.and Dixon, J.E. Ann. Biochem. 192, 262-267 (1991) SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: The Johns Hopkins (ii) TITLE OF INVENTION: Huntingtin-Asβociated Protein (iii) NUMBER OF SEQUENCES: 9 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: Banner & Witcoff, LTD. (B) STREET: 1001 G Street, NW suite 1100 (C) CITY: Washington (D) STATE: DC (E) COUNTRY: USA (F) ZIP: 20001 (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: (B) FILING DATE: (C) CLASSIFICATION: (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Kagan, Sarah A (B) REGISTRATION NUMBER: 32141 (C) REFERENCE/DOCKET NUMBER: 1107.01730 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: 202.508.9100 (B) TELEFAX: 202.508.9299 (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3144 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapient. (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: Met Ala Thr Leu Glu Lys Leu Met Lys Ala Phe Glu Ser Leu Lys Ser 1 5 10 15 Phe Gin Gin Gin Gin Gin Gin Gin Gin Gin Gin Gin Gin Gin Gin Gin 20 25 30 Gin Gin Gin Gin Gin Gin Gin Gin Pro Pro Pro Pro Pro Pro Pro Pro 35 40 45 Pro Pro Pro Gin Leu Pro Gin Pro Pro Pro Gin Ala Gin Pro Leu Leu 50 55 60 Pro Gin Pro Gin Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Gly Pro 65 70 75 80 Ala Val Ala Glu Glu Pro Leu His Arg Pro Lys Lys Glu Leu Ser Ala 85 90 95 Thr Lys Lys Asp Arg Val Aβn His Cys Leu Thr Ile Cys Glu Asn Ile 100 105 110 Val Ala Gin Ser Val Arg Asn Ser Pro Glu Phe Gin Lys Leu Leu Gly 115 120 125 Ile Ala Met Glu Leu Phe Leu Leu Cys Ser Asp Asp Ala Glu Ser Asp 130 135 140 Val Arg Met Val Ala Asp Glu Cys Leu Asn Lys Val Ile Lys Ala Leu 145 150 155 160 Met Asp Ser Asn Leu Pro Arg Leu Gin Leu Glu Leu Tyr Lys Glu Ile 165 170 175 Lys Lys Asn Gly Ala Pro Arg Ser Leu Arg Ala Ala Leu Trp Arg Phe 180 185 190 Ala Glu Leu Ala His Leu Val Arg Pro Gin Lys Cys Arg Pro Tyr Leu 195 200 205 Val Asn Leu Leu Pro Cys Leu Thr Arg Thr Ser Lys Arg Pro Glu Glu 210 215 220 Ser Val Gin Glu Thr Leu Ala Ala Ala Val Pro Lys Ile Met Ala Ser 225 230 235 240 Phe Gly Asn Phe Ala Asn Asp Asn Glu Ile Lys Val Leu Leu Lys Ala 245 250 255 Phe lie Ala Asn Leu Lys Ser Ser Ser Pro Thr Ile Arg Arg Thr Ala 260 265 270 Ala Gly ser Ala Val Ser Ile Cys Gin His Ser Arg Arg Thr Gin Tyr 275 280 285 Phe Tyr Ser Trp Leu Leu Asn Val Leu Leu Gly Leu Leu Val Pro Val 290 295 300 Glu Asp Glu His Ser Thr Leu Leu Ile Leu Gly Val Leu Leu Thr Leu 305 310 315 320 Arg Tyr Leu Val Pro Leu Leu Gin Gin Gin Val Lys Asp Thr Ser Leu 325 330 335 Lys Gly Ser Phe Gly Val Thr Arg Lys Glu Met Glu Val Ser Pro Ser 340 345 350 Ala Glu Gin Leu Val Gin Val Tyr Glu Leu Thr Leu His His Thr Gin 355 360 365 His Gin Asp His Asn Val Val Thr Gly Ala Leu Glu Leu Leu Gin Gin 370 375 380 Leu Phe Arg Thr Pro Pro Pro Glu Leu Leu Gin Thr Leu Thr Ala Val 385 390 395 400 Gly Gly Ile Gly Gin Leu Thr Ala Ala Lys Glu Glu Ser Gly Gly Arg 405 410 415 Ser Arg Ser Gly Ser Ile Val Glu Leu Ile Ala Gly Gly Gly Ser Ser 420 425 430 Cys Ser Pro Val Leu Ser Arg Lys Gin Lys Gly Lys Val Leu Leu Gly 435 440 445 Glu Glu Glu Ala Leu Glu Asp Asp Ser Glu Ser Arg Ser Asp Val Ser 450 455 460 Ser Ser Ala Leu Thr Ala Ser Val Lys Asp Glu Ile Ser Gly Glu Leu 465 470 475 480 Ala Ala Ser Ser Gly Val Ser Thr Pro Gly Ser Ala Gly His Asp Ile 485 490 495 Ile Thr Glu Gin Pro Arg Ser Gin His Thr Leu Gin Ala Asp Ser Val 500 505 510 Asp Leu Ala Ser Cys Asp Leu Thr Ser Ser Ala Thr Asp Gly Asp Glu 515 520 525 Glu Asp Ile Leu Ser His Ser Ser Ser Gin Val Ser Ala Val Pro Ser 530 535 540 Asp Pro Ala Met Asp Leu Asn Asp Gly Thr Gin Ala Ser Ser Pro Ile 545 550 555 560 Ser Asp Ser Ser Gin Thr Thr Thr Glu Gly Pro Asp Ser Ala Val Thr 565 570 575 Pro Ser Asp Ser Ser Glu Ile Val Leu Asp Gly Thr Asp Asn Gin Tyr 580 585 590 Leu Gly Leu Gin Ile Gly Gin Pro Gin Asp Glu Asp Glu Glu Ala Thr 595 600 605 Gly Ile Leu Pro Asp Glu Ala Ser Glu Ala Phe Arg Asn Ser Ser Met 610 615 620 Ala Leu Gin Gin Ala His Leu Leu Lys Asn Met Ser His Cys Arg Gin 625 630 635 640 Pro Ser Asp Ser Ser Val Asp Lys Phe Val Leu Arg Asp Glu Ala Thr 645 650 655 Glu Pro Gly Asp Gin Glu Asn Lys Pro Cys Arg Ile Lys Gly Asp Ile 660 665 670 Gly Gin Ser Thr Asp Asp Asp Ser Ala Pro Leu Val Hie Cys Val Arg 675 680 685 Leu Leu Ser Ala Ser Phe Leu Leu Thr Gly Gly Lys Asn Val Leu Val 690 695 700 Pro Asp Arg Asp Val Arg Val Ser Val Lys Ala Leu Ala Leu Ser Cys 705 710 715 720 Val Gly Ala Ala Val Ala Leu His Pro Glu Ser Phe Phe Ser Lys Leu 725 730 735 Tyr Lys Val Pro Leu Asp Thr Thr Glu Tyr Pro Glu Glu Gin Tyr Val 740 745 750 Ser Asp Ile Leu Asn Tyr Ile Asp His Gly Asp Pro Gin Val Arg Gly 755 760 765 Ala Thr Ala Ile Leu Cys Gly Thr Leu Ile Cys Ser Ile Leu Ser Arg 770 775 780 Ser Arg Phe His Val Gly Asp Trp Met Gly Thr Ile Arg Thr Leu Thr 785 790 795 800 Gly Asn Thr Phe Ser Leu Ala Asp Cys Ile Pro Leu Leu Arg Lys Thr 805 810 815 Leu Lys Asp Glu Ser Ser Val Thr Cys Lys Leu Ala Cys Thr Ala Val 820 825 830 Arg Asn Cys Val Met Ser Leu Cys Ser Ser Ser Tyr Ser Glu Leu Gly 835 840 845 Leu Gin Leu Ile Ile Asp Val Leu Thr Leu Arg Asn Ser Ser Tyr Trp 850 855 860 Leu Val Arg Thr Glu Leu Leu Glu Thr Leu Ala Glu Ile Asp Phe Arg 865 870 875 880 Leu Val Ser Phe Leu Glu Ala Lys Ala Glu Asn Leu His Arg Gly Ala 885 890 895 His His Tyr Thr Gly Leu Leu Lys Leu Gin Glu Arg Val Leu Asn Asn 900 905 910 Val Val Ile His Leu Leu Gly Asp Glu Asp Pro Arg Val Arg His Val 915 920 925 Ala Ala Ala Ser Leu Ile Arg Leu Val Pro Lys Leu Phe Tyr Lys Cys 930 935 940 Asp Gin Gly Gin Ala Asp Pro Val Val Ala Val Ala Arg Asp Gin Ser 945 950 955 960 Ser Val Tyr Leu Lys Leu Leu Met His Glu Thr Gin Pro Pro Ser His 965 970 975 Phe Ser Val Ser Thr Ile Thr Arg Ile Tyr Arg Gly Tyr Asn Leu Leu 980 985 990 Pro Ser Ile Thr Asp Val Thr Met Glu Asn Asn Leu Ser Arg Val Ile 995 1000 1005 Ala Ala Val Ser His Glu Leu Ile Thr Ser Thr Thr Arg Ala Leu Thr 1010 1015 1020 Phe Gly Cys Cys Glu Ala Leu Cys Leu Leu Ser Thr Ala Phe Pro Val 1025 1030 1035 1040 Cys He Trp Ser Leu Gly Trp His Cyβ Gly Val Pro Pro Leu Ser Ala 1045 1050 1055 Ser Asp Glu Ser Arg Lys Ser Cys Thr Val Gly Met Ala Thr Met Ile 1060 1065 1070 Leu Thr Leu Leu Ser Ser Ala Trp Phe Pro Leu Asp Leu Ser Ala His 1075 1080 1085 Gin Asp Ala Leu Ile Leu Ala Gly Asn Leu Leu Ala Ala Ser Ala Pro 1090 1095 1100 Lye Ser Leu Arg Ser Ser Trp Ala Ser Glu Glu Glu Ala Asn Pro Ala 1105 1110 1115 1120 Ala Thr Lys Gin Glu Glu Val Trp Pro Ala Leu Gly Asp Arg Ala Leu 1125 1130 1135 Val Pro Met Val Glu Gin Leu Phe Ser His Leu Leu Lys Val Ile Aβn 1140 1145 1150 Ile Cys Ala His Val Leu Asp Asp Val Ala Pro Gly Pro Ala Ile Lys 1155 1160 1165 Ala Ala Leu Pro Ser Leu Thr Asn Pro Pro Ser Leu Ser Pro Ile Arg 1170 1175 1180 Arg Lys Gly Lys Glu Lys Glu Pro Gly Glu Gin Ala Ser Val Pro Leu 1185 1190 1195 1200 Ser Pro Lys Lys Gly Ser Glu Ala Ser Ala Ala Ser Arg Gin Ser Asp 1205 1210 1215 Thr Ser Gly Pro Val Thr Thr Ser Lys Ser Ser Ser Leu Gly Ser Phe 1220 1225 1230 Tyr His Leu Pro Ser Tyr Leu Lys Leu His Asp Val Leu Lys Ala Thr 1235 1240 1245 His Ala Asn Tyr Lys Val Thr Leu Asp Leu Gin Asn Ser Thr Glu Lys 1250 1255 1260 Phe Gly Gly Phe Leu Arg Ser Ala Leu Asp Val Leu Ser Gin Ile Leu 1265 1270 1275 1280 Glu Leu Ala Thr Leu Gin Asp Ile Gly Lys Cys Val Glu Glu Ile Leu 1285 1290 1295 Gly Tyr Leu Lys Ser Cys Phe Ser Arg Glu Pro Met Met Ala Thr Val 1300 1305 1310 Cys Val Gin Gin Leu Leu Lys Thr Leu Phe Gly Thr Asn Leu Ala Ser 1315 1320 1325 Gin Phe Asp Gly Leu Ser Ser Asn Pro Ser Lys Ser Gin Gly Arg Ala 1330 1335 1340 Gin Arg Leu Gly Ser Ser Ser Val Arg Pro Gly Leu Tyr His Tyr Cys 1345 1350 1355 1360 Phe Met Ala Pro Tyr Thr His Phe Thr Gin Ala Leu Ala Asp Ala Ser 1365 1370 1375 Leu Arg Asn Met Val Gin Ala Glu Gin Glu Asn Asp Thr Ser Gly Trp 1380 1385 1390 Phe Asp Val Leu Gin Lys Val Ser Thr Gin Leu Lys Thr Asn Leu Thr 1395 1400 1405 Ser Val Thr Lys Asn Arg Ala Asp Lys Asn Ala Ile His Asn His Ile 1410 1415 1420 Arg Leu Phe Glu Pro Leu Val Ile Lys Ala Leu Lys Gin Tyr Thr Thr 1425 1430 1435 1440 Thr Thr Cys Val Gin Leu Gin Lys Gin Val Leu Asp Leu Leu Ala Gin 1445 1450 1455 Leu Val Gin Leu Arg Val Asn Tyr Cys Leu Leu Asp Ser Asp Gin Val 1460 1465 1470 Phe Ile Gly Phe Val Leu Lys Gin Phe Glu Tyr Ile Glu Val Gly Gin 1475 1480 1485 Phe Arg Glu Ser Glu Ala Ile Ile Pro Asn Ile Phe Phe Phe Leu Val 1490 1495 1500 Leu Leu Ser Tyr Glu Arg Tyr His Ser Lys Gin ie Ile Gly Ile Pro 1505 1510 1515 1520 Lys Ile Ile Gin Leu Cys Asp Gly Ile Met Ala Ser Gly Arg Lys Ala 1525 1530 1535 Val Thr His Ala Ile Pro Ala Leu Gin Pro Ile Val His Asp Leu Phe 1540 1545 1550 Val Leu Arg Gly Thr Asn Lys Ala Asp Ala Gly Lys Glu Leu Glu Thr 1555 1560 1565 Gin Lys Glu Val Val Val Ser Met Leu Leu Arg Leu Ile Gin Tyr His 1570 1575 1580 Gin Val Leu Glu Met Phe Ile Leu Val Leu Gin Gin Cys His Lys Glu 1585 1590 1595 1600 Asn Glu Asp Lys Trp Lys Arg Leu Ser Arg Gin Ile Ala Asp Ile Ile 1605 1610 1615 Leu Pro Met Leu Ala Lys Gin Gin Met His lie Asp Ser His Glu Ala 1620 1625 1630 Leu Gly Val Leu Asn Thr Leu Phe Glu Ile Leu Ala Pro Ser Ser Leu 1635 1640 1645 Arg Pro Val Asp Met Leu Leu Arg Ser Met Phe Val Thr Pro Asn Thr 1650 1655 1660 Met Ala Ser Val Ser Thr Val Gin Leu Trp Ile Ser Gly Ile Leu Ala 1665 1670 1675 1680 Ile Leu Arg Val Leu Ile Ser Gin Ser Thr Glu Asp Ile val Leu Ser 1685 1690 1695 Arg Ile Gin Glu Leu Ser Phe Ser Pro Tyr Leu Ile Ser Cys Thr Val 1700 1705 1710 lie Asn Arg Leu Arg Asp Gly Asp Ser Thr Ser Thr Leu Glu Glu His 1715 1720 1725 Ser Glu Gly Lys Gin Ile Lys Asn Leu Pro Glu Glu Thr Phe Ser Arg 1730 1735 1740 Phe Leu Leu Gin Leu Val Gly Ile Leu Leu Glu Asp Ile Val Thr Lys 1745 1750 1755 1760 Gin Leu Lys Val Glu Met Ser Glu Gin Gin His Thr Phe Tyr Cys Gin 1765 1770 1775 Glu Leu Gly Thr Leu Leu Met Cys Leu Ile His Ile Phe Lys Ser Gly 1780 1785 1790 Met Phe Arg Arg Ile Thr Ala Ala Ala Thr Arg Leu Phe Arg Ser Asp 1795 1800 1805 Gly Cys Gly Gly Ser Phe Tyr Thr Leu Asp Ser Leu Asn Leu Arg Ala 1810 1815 1820 Arg Ser Met Ile Thr Thr His Pro Ala Leu Val Leu Leu Trp Cys Gin 1825 1830 1835 1840 Ile Leu Leu Leu Val Asn His Thr Asp Tyr Arg Trp Trp Ala Glu Val 1845 1850 1855 Gin Gin Thr Pro Lys Arg His Ser Leu Ser Ser Thr Lys Leu Leu Ser 1860 1865 1870 Pro Gin Met Ser Gly Glu Glu Glu Asp Ser Asp Leu Ala Ala Lys Leu 1875 1880 1885 Gly Met Cys Asn Arg Glu Ile Val Arg Arg Gly Ala Leu Ile Leu Phe 1890 1895 1900 Cys Asp Tyr Val Cys Gin Asn Leu His Asp Ser Glu His Leu Thr Trp 1905 1910 1915 1920 Leu Ile Val Asn His Ile Gin Asp Leu Ile Ser Leu Ser His Glu Pro 1925 1930 1935 Pro Val Gin Asp Phe Ile Ser Ala Val His Arg Asn Ser Ala Ala Ser 1940 1945 1950 Gly Leu Phe Ile Gin Ala Ile Gin Ser Arg Cys Glu Asn Leu Ser Thr 1955 1960 1965 Pro Thr Met Leu Lys Lys Thr Leu Gin Cys Leu Glu Gly Ile His Leu 1970 1975 1980 Ser Gin Ser Gly Ala Val Leu Thr Leu Tyr Val Asp Arg Leu Leu Cys 1985 1990 1995 2000 Thr Pro Phe Arg Val Leu Ala Arg Met Val Asp Ile Leu Ala Cys Arg 2005 2010 2015 Arg Val Glu Met Leu Leu Ala Ala Asn Leu Gin Ser Ser Met Ala Gin 2020 2025 2030 Leu Pro Met Glu Glu Leu Asn Arg Ile Gin Glu Tyr Leu Gin Ser Ser 2035 2040 2045 Gly Leu Ala Gin Arg His Gin Arg Leu Tyr Ser Leu Leu Asp Arg Phe 2050 2055 2060 Arg Leu Ser Thr Met Gin Asp Ser Leu Ser Pro Ser Pro Pro Val Ser 2065 2070 2075 2080 Ser His Pro Leu Asp Gly Asp Gly His Val Ser Leu Glu Thr Val Ser 2085 2090 2095 Pro Asp Lys Asp Trp Tyr Val His Leu Val Lys Ser Gin Cys Trp Thr 2100 2105 2110 Arg Ser Asp Ser Ala Leu Leu Glu Gly Ala Glu Leu Val Asn Arg Ile 2115 2120 2125 Pro Ala Glu Asp Met Asn Ala Phe Met Met Asn Ser Glu Phe Aβn Leu 2130 2135 2140 Ser Leu Leu Ala Pro Cys Leu Ser Leu Gly Met Ser Glu Ile Ser Gly 2145 2150 2155 2160 Gly Gin Lys Ser Ala Leu Phe Glu Ala Ala Arg Glu Val Thr Leu Ala 2165 2170 2175 Arg Val Ser Gly Thr Val Gin Gin Leu Pro Ala Val His His Val Phe 2180 2185 2190 Gin Pro Glu Leu Pro Ala Glu Pro Ala Ala Tyr Trp Ser Lys Leu Asn 2195 2200 2205 Asp Leu Phe Gly Asp Ala Ala Leu Tyr Gin Ser Leu Pro Thr Leu Ala 2210 2215 2220 Arg Ala Leu Ala Gin Tyr Leu Val Val Val Ser Lys Leu Pro Ser His 2225 2230 2235 2240 Leu His Leu Pro Pro Glu Lys Glu Lys Asp Ile Val Lye Phe Val Val 2245 2250 2255 Ala Thr Leu Glu Ala Leu Ser Trp His Leu Ile His Glu Gin Ile Pro 2260 2265 2270 Leu Ser Leu Asp Leu Gin Ala Gly Leu Asp Cys Cys Cys Leu Ala Leu 2275 2280 2285 Gin Leu Pro Gly Leu Trp Ser Val Val Ser Ser Thr Glu Phe Val Thr 2290 2295 2300 His Ala Cys Ser Leu Ile Tyr Cys Val His Phe Ile Leu Glu Ala Val 2305 2310 2315 2320 Ala Val Gin Pro Gly Glu Gin Leu Leu Ser Pro Glu Arg Arg Thr Asn 2325 2330 2335 Thr Pro Lys Ala Ile Ser Glu Glu Glu Glu Glu Val Asp Pro Aβn Thr 2340 2345 2350 Gin Asn Pro Lys Tyr Ile Thr Ala Ala Cys Glu Met Val Ala Glu Met 2355 2360 2365 Val Glu Ser Leu Gin Ser Val Leu Ala Leu Gly His Lys Arg Asn Ser 2370 2375 2380 Gly Val Pro Ala Phe Leu Thr Pro Leu Leu Arg Asn Ile Ile Ile Ser 2385 2390 2395 2400 Leu Ala Arg Leu Pro Leu Val Asn Ser Tyr Thr Arg Val Pro Pro Leu 2405 2410 2415 Val Trp Lys Leu Gly Trp Ser Pro Lys Pro Gly Gly Asp Phe Gly Thr 2420 2425 2430 Ala Phe Pro Glu Ile Pro Val Glu Phe Leu Gin Glu Lys Glu Val Phe 2435 2440 2445 Lys Glu Phe Ile Tyr Arg Ile Asn Thr Leu Gly Trp Thr Ser Arg Thr 2450 2455 2460 Gin Phe Glu Glu Thr Trp Ala Thr Leu Leu Gly Val Leu Val Thr Gin 2465 2470 2475 2480 Pro Leu Val Met Glu Gin Glu Glu Ser Pro Pro Glu Glu Asp Thr Glu 2485 2490 2495 Arg Thr Gin Ile Asn Val Leu Ala Val Gin Ala Ile Thr Ser Leu Val 2500 2505 2510 Leu Ser Ala Met Thr Val Pro Val Ala Gly Asn Pro Ala Val Ser Cys 2515 2520 2525 Leu Glu Gin Gin Pro Arg Asn Lys Pro Leu Lys Ala Leu Asp Thr Arg 2530 2535 2540 Phe Gly Arg Lys Leu Ser Ile Ile Arg Gly Ile Val Glu Gin Glu Ile 2545 2550 2555 2560 Gin Ala Met Val Ser Lys Arg Glu Asn Ile Ala Thr His His Leu Tyr 2565 2570 2575 Gin Ala Trp Asp Pro Val Pro Ser Leu Ser Pro Ala Thr Thr Gly Ala 2580 2585 2590 Leu Ile Ser His Glu Lys Leu Leu Leu Gin Ile Asn Pro Glu Arg Glu 2595 2600 2605 Leu Gly Ser Met Ser Tyr Lys Leu Gly Gin Val Ser Ile His Ser Val 2610 2615 2620 Trp Leu Gly Asn Ser Ile Thr Pro Leu Arg Glu Glu Glu Trp Asp Glu 2625 2630 2635 2640 Glu Glu Glu Glu Glu Ala Asp Ala Pro Ala Pro Ser Ser Pro Pro Thr 2645 2650 2655 Ser Pro Val Asn Ser Arg Lys His Arg Ala Gly Val Asp Ile His Ser 2660 2665 2670 Cys Ser Gin Phe Leu Leu Glu Leu Tyr Ser Arg Trp Ile Leu Pro Ser 2675 2680 2685 Ser Ser Ala Arg Arg Thr Pro Ala Ile Leu Ile Ser Glu Val Val Arg 2690 2695 2700 Ser Leu Leu Val Val Ser Asp Leu Phe Thr Glu Arg Asn Gin Phe Glu 2705 2710 2715 2720 Leu Met Tyr Val Thr Leu Thr Glu Leu Arg Arg Val His Pro Ser Glu 2725 2730 2735 Asp Glu Ile Leu Ala Gin Tyr Leu Val Pro Ala Thr Cys Lys Ala Ala 2740 2745 2750 Ala Val Leu Gly Met Asp Lys Ala Val Ala Glu Pro Val Ser Arg Leu 2755 2760 2765 Leu Glu Ser Thr Leu Arg Ser Ser His Leu Pro Ser Arg Val Gly Ala 2770 2775 2780 Leu His Gly Val Leu Tyr Val Leu Glu Cys Asp Leu Leu Asp Asp Thr 2785 2790 2795 2800 Ala Lye Gin Leu lie Pro Val Ile Ser Asp Tyr Leu Leu Ser Asn Leu 2805 2810 2815 Lys Gly Ile Ala His Cys Val Asn Ile His Ser Gin Gin His Val Leu 2820 2825 2830 Val Met Cys Ala Thr Ala Phe Tyr Leu lie Glu Asn Tyr Pro Leu Asp 2835 2840 2845 Val Gly Pro Glu Phe Ser Ala Ser Ile Ile Gin Met Cys Gly Val Met 2850 2855 2860 Leu Ser Gly Ser Glu Glu Ser Thr Pro Ser Ile Ile Tyr His Cys Ala 2865 2870 2875 2880 Leu Arg Gly Leu Glu Arg Leu Leu Leu Ser Glu Gin Leu Ser Arg Leu 2885 2890 2895 Asp Ala Glu Ser Leu Val Lys Leu Ser Val Asp Arg Val Asn Val His 2900 2905 2910 Ser Pro His Arg Ala Met Ala Ala Leu Gly Leu Met Leu Thr Cys Met 2915 2920 2925 Tyr Thr Gly Lys Glu Lys Val Ser Pro Gly Arg Thr Ser Asp Pro Asn 2930 2935 2940 Pro Ala Ala Pro Asp Ser Glu Ser Val Ile Val Ala Met Glu Arg Val 2945 2950 2955 2960 Ser Val Leu Phe Asp Arg Ile Arg Lys Gly Phe Pro Cys Glu Ala Arg 2965 2970 2975 Val Val Ala Arg Ile Leu Pro Gin Phe Leu Asp Asp Phe Phe Pro Pro 2980 2985 2990 Gin Asp Ile Met Asn Lys Val Ile Gly Glu Phe Leu Ser Aβn Gin Gin 2995 3000 3005 Pro Tyr Pro Gin Phe Met Ala Thr Val Val Tyr Lys Val Phe Gin Thr 3010 3015 3020 Leu His Ser Thr Gly Gin Ser Ser Met Val Arg Asp Trp Val Met Leu 3025 3030 3035 3040 Ser Leu Ser Asn Phe Thr Gin Arg Ala Pro Val Ala Met Ala Thr Trp 3045 3050 3055 Ser Leu Ser Cys Phe Phe Val Ser Ala Ser Thr Ser Pro Trp Val Ala 3060 3065 3070 Ala Ile Leu Pro His Val Ile Ser Arg Met Gly Lys Leu Glu Gin Val 3075 3080 3085 Asp Val Asn Leu Phe Cys Leu Val Ala Thr Asp Phe Tyr Arg His Gin 3090 3095 3100 lie Glu Glu Glu Leu Asp Arg Arg Ala Phe Gin Ser Val Leu Glu Val 3105 3110 3115 3120 Val Ala Ala Pro Gly Ser Pro Tyr His Arg Leu Leu Thr Cys Leu Arg 3125 3130 3135 Aβn Val His Lys Val Thr Thr Cys 3140 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 960 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: CTCTACTCAG ATTCTGATGA GGAGGATGAG GATGAAGAAG AGGAGGAGGA AGAAAAGGAG 60 GCAGAAGAGG AACAGGAAGA AG AGAAGCA GAGGAAGACC TGCAGTGTGC TCATCCCTGT 120 GATGCCCCTA AGCTGATTTC GCAGGAGGCA TTGCTGCACC AGCACCACTG CCCACAGCTG 180 GAAGCCTTGC AGGAGAAGCT GAGGCTGCTG GAGGAGGAGA ATCATCAGCT GAGAGAAGAG 240 GCCTCTCAAC TCGACACTCT TGAGGATGAG GAACAGATGC TCATTCTGGA GTGTGTGGAG 300 CAGTTTTCGG AGGCCAGCCA ACAGATGGCT GAGCTGTCGG AGGTGCTGGT GCTCAGGCTG 360 GAAAACTATG AACGGCAGCA GCAGGAGGTC GCTCGGCTGC AGGCCCAGGT GCTGAAGCTG 420 CAGCAGCGCT GCCGGATGTA TGGGGCTGAG ACTGAAAAGT TGCAGAAGCA GCTGGCTTCG 480 GAGAAGGAAA TCCAGATGCA GCTCCAGGAA GAGGAGACTC TGCCTGGTTT CCAGGAGACG 540 CTGGCTGAGG AGCTCAGAAC GTCTCTAAGG AGGATGATCT CAGACCCTGT GTATTTTATG 600 GAGAGGAATT ATGAGATGCC CAGAGGGGAC ACATCCAGCC TAAGGTATGA TTTTCGCTAC 660 AGTGAGGATC GAGAGCAGGT GCGGGGGTTT GAGGCTGAGG AAGGGTTGAT GCTGGCAGCG 720 GATATCATGC GGGGGGAAGA TTTCACGCCT GCGGAGGAGT TGGTGCCCCA GGAGGAGCTG 780 GGGGCTGCCA AGAAGGTGCC GGCTGAGGAA GGGGTGATGG AAGAGGCAGA GCTGGTGTCA 840 GAGGAGACCG AGGGCTGGGA GGAGGTGGAA CTGGAGCTGG ATGAGGCAAC GCGGATGAAC 900 GTGGTGACAT CAACCCTGGA GGCCAGCGGC TTGGGCCCTT CACACCTGGA CATGAATTAT 960 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3708 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Rattus norvegicus (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: GGAATTCGGC ACGAGGCGAC AGCGGACTGC AGAGTCAAGA TGCGCCCGAA GGACCAGGTG 60 CAGAGCAGTG CCGGGGACGG GACGGGGTCG GGGGACCCAG CAACAGGCAC CCCCACGACC 120 CAGCCTGCAG CGGATCCCGC TCCGGAGCCC TCGGCAGAGC CCAAACCTGC TCCGGCGCAG 180 GGAACCGGGT CCGGACAGAA ATCAGGATCC CGAACCAAGA CAGGAGGAAG CTTTTGTAGG 240 TCCAGGATCC GTGGCGACTC GGACGCACCA TGGACCCGCT ACATATTCCA GGGGCCTTAC 300 GGTCCCCGGG CTACTGGCCT GGGCACTGGA AGGGCTGAGG GAATCTGGAA GACGCCAGCC 360 GCGTACATCG GCCGAAGGCC CGGCGTGTCC GGCCCTGAGC GCGCGGCGTT TATTCGAGAG 420 CTGCAGGAAG CGCTGTGTCC TAATCCACTG CCCAGGAAGA AGATCACCGA AGATGATATC 480 AAAGTGATGT TGTATTTGCT GGAAGAGAAA GAACGGGACC TGAACACAGC CGCTCGCATC 540 GGCCAGTCCC TGGTGAAACA GAATAGTGTT CTGATGGAGG AGAATAACAA GCTGGAAACC 600 ATGCTGGGCT CAGCCAGAGA GGAGATTTTA CATCTCCGGA AGCAGGTGAA CCTGCGAGAT 660 GATCTCCTTC AGCTCTACTC GGACTCCGAT GACGATGAGG AGGATGAAGA GGATGAGGAA 720 GAGGAAGAGG GAGAAGAGGA GGAACGAGAA GGACAGAGGG ACCAAGATCA GCAGCACGAC 780 CATCCCTATG GTGCCCCCAA GCCGCCCCCT AAGGCTGAGA CGCTGCACCA CTGCCCACAG 840 CTGGAAGCCC TGAAGCAGAA GCTGAAACTG CTGGAAGAAG AGAACGACCA TCTTCGAGAG 900 GAGGCCTCCC ACCTTGACAA CCTGGAAGAC AAAGAACAGA TGCTCATTCT GGAGTGTGTG 960 GAACAGTTTT CTGAAGCCAG CCAGCAGATG GCAGAGCTAT CCGAGGTGTT GGTGCTGAGG 1020 CTGGAAGGCT ATGAGAGGCA GCAGAAGGAG ATCACTCAGC TGCAGGCCGA GATCACCAAG 1080 CTACAACAGC GTTGTCAGTC TTATGGGGCC CAGACGGAGA AACTGCAGCA GCAGCTGGCC 1140 TCAGAGAAGG GAGTCCACCC AGAGAGCCTG CGAGCTGGCT CCCACATGCA GGATTATGGA 1200 AGCAGGCCTC GTGAACGCCA GGAGGATGGG AAGAGCCATC GTCAGCGTTC CTCAATGCCT 1260 GCAGGTTCTG TCACCCACTA TGGATACAGT GTGCCTCTGG ATGCACTTCC AAGTTTCCCA 1320 GAGACACTGG CGGAGGAGCT CCGGACATCC CTGAGGAAGT TCATCACTGA CCCTGCGTAT 1380 TTCATGGAGA GATGTGACAC TCGCTGCAGA GAGGAACGAA AGAAGGAGCA GGGGACAATG 1440 CCACCCCCAC CGGTGCAAGA TCTCAAGCCG CCTGAAGATT TCGAGGCTCC AGAGGAGCTG 1500 GTTCCTGAGG AGGAGCTGGG GGCCATAGAA GAGGTGGGGA CAGCTGAGGA TGGGCCGGCA 1560 GAAGAGACAG AGCAGGCATC TGAGGAGACC GAGGCCTGGG AGGAGGTGGA ACCGGAGGTG 1620 GACGAGGCCA CAAGGATGAA TGTGGTGGTC TCTGCCCTGG AGGCCAGCGG CCTGGGCCCT 1680 TCACACCTGG ACATGAAGTA TGTCCTCCAG CAACTGTCCA ACTGGCAGGA CGCCCATTCT 1740 AAGCGGCAGC AGAAGCAGAA GGTGGTCCCG AAAGGTGAGT GTTCCCGCAG AGGACACCCT 1800 CCTGCCAGTG GGACAAGCTA CCGATCATCA ACCCTATGAG AGGTGAGAGT AGGTGAGACC 1860 CCCCACCCCC AAGGGCTCAC TTACCTCACC TTGGTCCCAC TCGGTGTGCT GATTTGCATG 1920 GACTTTGCAT ACTATTTGCA TAGTATTTAC ATACTTGCCT CCAGTCCCCC TTGGCTAGAA 1980 CTGCTGCCTC AGTGTTTATT TATGCAAAAT TTGCTTACAA GTCCAGCTAT CCATCCACCT 2040 TCTTCCTGGG GGGGCTGAAC TGGGAATCAG GGTTGGACTA TACACTCCTT TGTACCTCAA 2100 CTTCCTGTCT CTGCCCATCC TCTCCCTATC CCCAATTCCT CTACCCTCAA GACTCCCCAG 2160 CCCCGCAGCA GCAAACAAAC ATGGGGGGCG GGATCGTGGA GCAGCAGCCC ATAGTGCCGA 2220 CCCAGGACTC TCAGAGGCTG GAGGAGGACA GGGCCACTCA CTCTCCCAGT GCCAGGGAGG 2280 AAGAGGGGCC TTCTGGGGCC ACCTAGGCCT GGAACGAAGG CCTCTGCCAG CAAAGGCCAC 2340 AGTTGGACCA ATCCCCCGGG TGAGTGTGGG GCTCCCGCAG GGTGGTAGGG GCGGGTCAGG 2400 GTCTCCTCTG CCTATCTGGA AACCCCAGTT CCTAGGAGAT GCTGTCCGTG AAATCAGATG 2460 ATTTAGCATT GGATCTCTTT CTGTGTTAGC TCCTTTCGTC TTCTGGCCCT GTGGGTGGGT 2520 GGATGTGATT GTGCACCATT TATTAGAAGA GGAATCTGAA GTTCAGAGAT CTGAGATGAC 2580 TTGGCCAAGG TCATACAGCT AGGAGCAGAT CTGAGCCGGT GTCTCTTGAG GACTGAAACC 2640 CTGGCCTGGT CTCACTGCCG TCCACACCTC CTCCTGCTCC CGTCCTTGAG CTCCACCTGA 2700 AGCTTTGGCA GCCTCCCTCA TGTCGGTCTG TTGCCATTTT TGGGGGGCAC CAGTGAGTGG 2760 AACATCACTG GCTGCAGCAC AGTCAAATCA TGCAGGTTGT AAGCTGAGTC AGCACACCCG 2820 TGGCTGACCA CGGGAGACCG GGAGACCTGA GTAACGGCTA GCGGTGGCTT TCTGTTGTCT 2880 CCTGACCCGG CTGACTGGGT TTGAGGGTGA TCTGTCTCCC GGTTCACCCC CTTCTCTTCC 2940 TGGCTTTGCT GTTTCTCAGC AGCCTGAAGA AGACCCTCAG ACAGCTCGAG ACCCCTCCTT 3000 CCACTGCCTG GCCAAGTCCG ACCCTTCCTT GCTTTCCTCT GAGACAGGAC CCCCACCCCC 3060 ATGTCTGTCA GCAGTTCTCC CTCTGTCTTG ATAGATCTTT CCCCTCTTGT AGGACAGGCT 3120 GAAAGAACCC CAGTCCCTCA TTCTGAAACT GGGGCCAAAG CTGTGCCTGC TGGAGCCCCA 3180 GGGGGTGGAA GCAACCTGTG GGTAGATTGG TTGGTTACCA GACCCAGCTC TGAGATGGTG 3240 TGGGTGCGCA CATCCCTAGG GTAGGCAGTT ATTGGGGGGA CCCTTTCTAT CCCTTGAACC 3300 TCTCACGTAA AGGGACTTCC CCAGTCCTGG TTGGCTTTTG GAACCTGGTC CTTCTTGCTG 3360 TTTTTTACCC TTCCCCGTTT CTATTACTGC GTGTAACGTA AAGTGTATCT GAGTGAGGGT 3420 GGTGGGAACC CTCTGTCCAG TGCTGTCTCT GTCTCCTATG GCCTGTGAGT TTCCTTCTAG 3480 AGTTCTACTC TTCTCCACCT CTTTGCTCAT ACAGAGCTGT GGCCTTGGCG CCTGCCCTGC 3540 TTCTGCAGTG CTTCATTGTC TCGTAGCTTG TCAGCTGACA CCGGAACCGC GGACGGGACG 3600 AAGACGTCAC GAAGTAACAG AGCATCGAAC AGTCGATTTG TATTGATGTA TGTGCCAATG 3660 TGGGAAATAA AGACCTTGTG AGATAAAAAA AAAAAAAAAA AAAAAAAA 3708 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 599 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (vi) ORIGINAL SOURCE: (A) ORGANISM: Rattus norvegicus (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: Met Arg Pro Lys Asp Gin Val Gin Ser Ser Ala Gly Asp Gly Thr Gly 1 5 10 15 Ser Gly Asp Pro Ala Thr Gly Thr Pro Thr Thr Gin Pro Ala Ala Asp 20 25 30 Pro Ala Pro Glu Pro Ser Ala Glu Pro Lys Pro Ala Pro Ala Gin Gly 35 40 45 Thr Gly Ser Gly Gin Lys Ser Gly Ser Arg Thr Lys Thr Gly Gly Ser 50 55 60 Phe Cys Arg Ser Arg Ile Arg Gly Asp Ser Asp Ala Pro Trp Thr Arg 65 70 75 80 Tyr Ile Phe Gin Gly Pro Tyr Gly Pro Arg Ala Thr Gly Leu Gly Thr 85 90 95 Gly Arg Ala Glu Gly Ile Trp Lys Thr Pro Ala Ala Tyr Ile Gly Arg 100 105 110 Arg Pro Gly Val Ser Gly Pro Glu Arg Ala Ala Phe Ile Arg Glu Leu 115 120 125 Gin Glu Ala Leu Cys Pro Asn Pro Leu Pro Arg Lys Lys Ile Thr Glu 130 135 140 Asp Asp Ile Lys Val Met Leu Tyr Leu Leu Glu Glu Lys Glu Arg Asp 145 150 155 160 Leu Aβn Thr Ala Ala Arg Ile Gly Gin Ser Leu Val Lye Gin Aβn Ser 165 170 175 Val Leu Met Glu Glu Asn Asn Lys Leu Glu Thr Met Leu Gly Ser Ala 180 185 190 Arg Glu Glu Ile Leu His Leu Arg Lys Gin Val Asn Leu Arg Asp Aβp 195 200 205 Leu Leu Gin Leu Tyr Ser Asp Ser Asp Asp Asp Glu Glu Aβp Glu Glu 210 215 220 Asp Glu Glu Glu Glu Glu Gly Glu Glu Glu Glu Arg Glu Gly Gin Arg 225 230 235 240 Asp Gin Asp Gin Gin His Asp His Pro Tyr Gly Ala Pro Lys Pro Pro 245 250 255 Pro Lys Ala Glu Thr Leu His His Cys Pro Gin Leu Glu Ala Leu Lys 260 265 270 Gin Lys Leu Lys Leu Leu Glu Glu Glu Asn Asp His Leu Arg Glu Glu 275 280 285 Ala Ser His Leu Asp Asn Leu Glu Asp Lys Glu Gin Met Leu Ile Leu 290 295 300 Glu Cys Val Glu Gin Phe Ser Glu Ala Ser Gin Gin Met Ala Glu Leu 305 310 315 320 Ser Glu Val Leu Val Leu Arg Leu Glu Gly Tyr Glu Arg Gin Gin Lys 325 330 335 Glu Ile Thr Gin Leu Gin Ala Glu Ile Thr Lys Leu Gin Gin Arg Cys 340 345 350 Gin Ser Tyr Gly Ala Gin Thr Glu Lye Leu Gin Gin Gin Leu Ala Ser 355 360 365 Glu Lys Gly Val His Pro Glu Ser Leu Arg Ala Gly Ser His Met Gin 370 375 380 Aβp Tyr Gly Ser Arg Pro Arg Glu Arg Gin Glu Asp Gly Lye Ser Hie 385 390 395 400 Arg Gin Arg Ser Ser Met Pro Ala Gly Ser Val Thr His Tyr Gly Tyr 405 410 415 Ser Val Pro Leu Asp Ala Leu Pro Ser Phe Pro Glu Thr Leu Ala Glu 420 425 430 Glu Leu Arg Thr Ser Leu Arg Lys Phe Ile Thr Asp Pro Ala Tyr Phe 435 440 445 Met Glu Arg Cys Asp Thr Arg Cys Arg Glu Glu Arg Lys Lye Glu Gin 450 455 460 Gly Thr Met Pro Pro Pro Pro Val Gin Asp Leu Lys Pro Pro Glu Aβp 465 470 475 480 Phe Glu Ala Pro Glu Glu Leu Val Pro Glu Glu Glu Leu Gly Ala Ile 485 490 495 Glu Glu Val Gly Thr Ala Glu Asp Gly Pro Ala Glu Glu Thr Glu Gin 500 505 510 Ala Ser Glu Glu Thr Glu Ala Trp Glu Glu Val Glu Pro Glu Val Asp 515 520 525 Glu Ala Thr Arg Met Asn Val Val Val Ser Ala Leu Glu Ala Ser Gly 530 535 540 Leu Gly Pro Ser His Leu Asp Met Lys Tyr Val Leu Gin Gin Leu Ser 545 550 555 560 Asn Trp Gin Asp Ala His Ser Lys Arg Gin Gin Lys Gin Lys Val Val 565 570 575 Pro Lys Gly Glu Cys Ser Arg Arg Gly His Pro Pro Ala Ser Gly Thr 580 585 590 Ser Tyr Arg Ser Ser Thr Leu 595 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3201 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Rattus norvegicus (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 40..1927 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: GGAATTCGGC ACGAGGCGAC AGCGGACTGC AGAGTCAAG ATG CGC CCG AAG GAC 54 Met Arg Pro Lys Asp 1 5 CAG GTG CAG AGC AGT GCC GGG GAC GGG ACG GGG TCG GGG GAC CCA GCA 102 Gin Val Gin Ser Ser Ala Gly Aβp Gly Thr Gly Ser Gly Aβp Pro Ala 10 15 20 ACA GGC ACC CCC ACG ACC CAG CCT GCA GCG GAT CCC GCT CCG GAG CCC 150 Thr Gly Thr Pro Thr Thr Gin Pro Ala Ala Asp Pro Ala Pro Glu Pro 25 30 35 TCG GCA GAG CCC AAA CCT GCT CCG GCG CAG GGA ACC GGG TCC GGA CAG 198 Ser Ala Glu Pro Lys Pro Ala Pro Ala Gin Gly Thr Gly Ser Gly Gin 40 45 50 AAA TCA GGA TCC CGA ACC AAG ACA GGA GGA AGC TTT TGT AGG TCC AGG 246 Lys Ser Gly Ser Arg Thr Lys Thr Gly Gly Ser Phe Cys Arg Ser Arg 55 60 65 ATC CGT GGC GAC TCG GAC GCA CCA TGG ACC CGC TAC ATA TTC CAG GGG 294 He Arg Gly Asp Ser Aβp Ala Pro Trp Thr Arg Tyr Ile Phe Gin Gly 70 75 80 85 CCT TAC GGT CCC CGG GCT ACT GGC CTG GGC ACT GGA AGG GCT GAG GGA 342 Pro Tyr Gly Pro Arg Ala Thr Gly Leu Gly Thr Gly Arg Ala Glu Gly 90 95 100 ATC TGG AAG ACG CCA GCC GCG TAC ATC GGC CGA AGG CCC GGC GTG TCC 390 Ile Trp Lye Thr Pro Ala Ala Tyr Ile Gly Arg Arg Pro Gly Val Ser 105 110 115 GGC CCT GAG CGC GCG GCG TTT ATT CGA GAG CTG CAG GAA GCG CTG TGT 438 Gly Pro Glu Arg Ala Ala Phe Ile Arg Glu Leu Gin Glu Ala Leu Cys 120 125 130 CCT AAT CCA CTG CCC AGG AAG AAG ATC ACC GAA GAT GAT ATC AAA GTG 486 Pro Asn Pro Leu Pro Arg Lys Lys Ile Thr Glu Asp Asp Ile Lys Val 135 140 145 ATG TTG TAT TTG CTG GAA GAG AAA GAA CGG GAC CTG AAC ACA GCC GCT 534 Met Leu Tyr Leu Leu Glu Glu Lys Glu Arg Asp Leu Asn Thr Ala Ala 150 155 160 165 CGC ATC GGC CAG TCC CTG GTG AAA CAG AAT AGT GTT CTG ATG GAG GAG 582 Arg Ile Gly Gin Ser Leu Val Lys Gin Asn Ser Val Leu Met Glu Glu 170 175 180 AAT AAC AAG CTG GAA ACC ATG CTG GGC TCA GCC AGA GAG GAG ATT TTA 630 Asn Asn Lys Leu Glu Thr Met Leu Gly Ser Ala Arg Glu Glu Ile Leu 185 190 195 CAT CTC CGG AAG CAG GTG AAC CTG CGA GAT GAT CTC CTT CAG CTC TAC 678 His Leu Arg Lys Gin Val Asn Leu Arg Asp Asp Leu Leu Gin Leu Tyr 200 205 210 TCG GAC TCC GAT GAC GAT GAG GAG GAT GAA GAG GAT GAG GAA GAG GAA 726 Ser Asp Ser Asp Asp Asp Glu Glu Asp Glu Glu Asp Glu Glu Glu Glu 215 220 225 GAG GGA GAA GAG GAG GAA CGA GAA GGA CAG AGG GAC CAA GAT CAG CAG 774 Glu Gly Glu Glu Glu Glu Arg Glu Gly Gin Arg Asp Gin Asp Gin Gin 230 235 240 245 CAC GAC CAT CCC TAT GGT GCC CCC AAG CCG CCC CCT AAG GCT GAG ACG 822 His Asp His Pro Tyr Gly Ala Pro Lys Pro Pro Pro Lys Ala Glu Thr 250 255 260 CTG CAC CAC TGC CCA CAG CTG GAA GCC CTG AAG CAG AAG CTG AAA CTG 870 Leu His His Cys Pro Gin Leu Glu Ala Leu Lys Gin Lys Leu Lys Leu 265 270 275 CTG GAA GAA GAG AAC GAC CAT CTT CGA GAG GAG GCC TCC CAC CTT GAC 918 Leu Glu Glu Glu Asn Asp His Leu Arg Glu Glu Ala Ser His Leu Asp 280 285 290 AAC CTG GAA GAC AAA GAA CAG ATG CTC ATT CTG GAG TGT GTG GAA CAG 966 Asn Leu Glu Asp Lys Glu Gin Met Leu Ile Leu Glu Cys Val Glu Gin 295 300 305 TTT TCT GAA GCC AGC CAG CAG ATG GCA GAG CTA TCC GAG GTG TTG GTG 1014 Phe Ser Glu Ala Ser Gin Gin Met Ala Glu Leu Ser Glu Val Leu Val 310 315 320 325 CTG AGG CTG GAA GGC TAT GAG AGG CAG CAG AAG GAG ATC ACT CAG CTG 1062 Leu Arg Leu Glu Gly Tyr Glu Arg Gin Gin Lys Glu Ile Thr Gin Leu 330 335 340 CAG GCC GAG ATC ACC AAG CTA CAA CAG CGT TGT CAG TCT TAT GGG GCC 1110 Gin Ala Glu Ile Thr Lys Leu Gin Gin Arg Cys Gin Ser Tyr Gly Ala 345 350 355 CAG ACG GAG AAA CTG CAG CAG CAG CTG GCC TCA GAG AAG GGA GTC CAC 1158 Gin Thr Glu Lys Leu Gin Gin Gin Leu Ala Ser Glu Lys Gly Val His 360 365 370 CCA GAG AGC CTG CGA GCT GGC TCC CAC ATG CAG GAT TAT GGA AGC AGG 1206 Pro Glu Ser Leu Arg Ala Gly Ser His Met Gin Asp Tyr Gly Ser Arg 375 380 385 CCT CGT GAA CGC CAG GAG GAT GGG AAG AGC CAT CGT CAG CGT TCC TCA 1254 Pro Arg Glu Arg Gin Glu Asp Gly Lys Ser His Arg Gin Arg Ser Ser 390 395 400 405 ATG CCT GCA GGT TCT GTC ACC CAC TAT GGA TAC AGT GTG CCT CTG GAT 1302 Met Pro Ala Gly Ser Val Thr His Tyr Gly Tyr Ser Val Pro Leu Asp 410 415 420 GCA CTT CCA AGT TTC CCA GAG ACA CTG GCG GAG GAG CTC CGG ACA TCC 1350 Ala Leu Pro Ser Phe Pro Glu Thr Leu Ala Glu Glu Leu Arg Thr Ser 425 430 435 CTG AGG AAG TTC ATC ACT GAC CCT GCG TAT TTC ATG GAG AGA TGT GAC 1398 Leu Arg Lys Phe Ile Thr Asp Pro Ala Tyr Phe Met Glu Arg Cys Asp 440 445 450 ACT CGC TGC AGA GAG GAA CGA AAG AAG GAG CAG GGG ACA ATG CCA CCC 1446 Thr Arg Cys Arg Glu Glu Arg Lys Lys Glu Gin Gly Thr Met Pro Pro 455 460 465 CCA CCG GTG CAA GAT CTC AAG CCG CCT GAA GAT TTC GAG GCT CCA GAG 1494 Pro Pro Val Gin Asp Leu Lys Pro Pro Glu Asp Phe Glu Ala Pro Glu 470 475 480 485 GAG CTG GTT CCT GAG GAG GAG CTG GGG GCC ATA GAA GAG GTG GGG ACA 1542 Glu Leu Val Pro Glu Glu Glu Leu Gly Ala Ile Glu Glu Val Gly Thr 490 495 500 GCT GAG GAT GGG CCG GCA GAA GAG ACA GAG CAG GCA TCT GAG GAG ACC 1590 Ala Glu Asp Gly Pro Ala Glu Glu Thr Glu Gin Ala Ser Glu Glu Thr 505 510 515 GAG GCC TGG GAG GAG GTG GAA CCG GAG GTG GAC GAG GCC ACA AGG ATG 1638 Glu Ala Trp Glu Glu Val Glu Pro Glu Val Asp Glu Ala Thr Arg Met 520 525 530 AAT GTG GTG GTC TCT GCC CTG GAG GCC AGC GGC CTG GGC CCT TCA CAC 1686 Asn Val Val Val Ser Ala Leu Glu Ala Ser Gly Leu Gly Pro Ser His 535 540 545 CTG GAC ATG AAG TAT GTC CTC CAG CAA CTG TCC AAC TGG CAG GAC GCC 1734 Leu Asp Met Lys Tyr Val Leu Gin Gin Leu Ser Asn Trp Gin Asp Ala 550 555 560 565 CAT TCT AAG CGG CAG CAG AAG CAG AAG GTG GTC CCG AAA GAC TCC CCA 1782 His Ser Lye Arg Gin Gin Lys Gin Lys Val Val Pro Lys Asp Ser Pro 570 575 580 GCC CCG CAG CAG CAA ACA AAC ATG GGG GGC GGG ATC GTG GAG CAG CAG 1830 Ala Pro Gin Gin Gin Thr Aβn Met Gly Gly Gly Ile Val Glu Gin Gin 585 590 595 CCC ATA GTG CCG ACC CAG GAC TCT CAG AGG CTG GAG GAG GAC AGG GCC 1878 Pro Ile Val Pro Thr Gin Asp Ser Gin Arg Leu Glu Glu Asp Arg Ala 600 605 610 ACT CAC TCT CCC AGT GCC AGG GAG GAA GAG GGG CCT TCT GGG GCC ACC T 1927 Thr His Ser Pro Ser Ala Arg Glu Glu Glu Gly Pro Ser Gly Ala Thr 615 620 625 AGGCCTGGAA CGAAGGCCTC TGCCAGCAAA GGCCACAGTT GGACCAATCC CCCGGCTCCT 1987 TTCGTCTTCT GGCCCTGTGG GTGGGTGGAT GTGATTGTGC ACCATTTATT AGAAGAGGAA 2047 TCTGAAGTTC AGAGATCTGA GATGACTTGG CCAAGGTCAT ACAGCTAGGA GCAGATCTGA 2107 GCCGGTGTCT CTTGAGGACT GAAACCCTGG CCTGGTCTCA CTGCCGTCCA CACCTCCTCC 2167 TGCTCCCGTC CTTGAGCTCC ACCTGAAGCT TTGGCAGCCT CCCTCATGTC GGTCTGTTGC 2227 CATTTTTGGG GGGCACCAGT GAGTGGAACA TCACTGGCTG CAGCACAGTC AAATCATGCA 2287 GGTTGTAAGC TGAGTCAGCA CACCCGTGGC TGACCACGGG AGACCGGGAG ACCTGAGTAA 2347 CGGCTAGCGG TGGCTTTCTG TTGTCTCCTG ACCCGGCTGA CTGGGTTTGA GGGTGATCTG 2407 TCTCCCGGTT CACCCCCTTC TCTTCCTGGC TTTGCTGTTT CTCAGCAGCC TGAAGAAGAC 2467 CCTCAGACAG CTCGAGACCC CTCCTTCCAC TGCCTGGCCA AGTCCGACCC TTCCTTGCTT 2527 TCCTCTGAGA CAGGACCCCC ACCCCCATGT CTGTCAGCAG TTCTCCCTCT GTCTTGATAG 2587 ATCTTTCCCC TCTTGTAGGA CAGGCTGAAA GAACCCCAGT CCCTCATTCT GAAΛCTGGGG 2647 CCAAAGCTGT GCCTGCTGGA GCCCCAGGGG GTGGAAGCAA CCTGTGGGTA GATTGGTTGG 2707 TTACCAGACC CAGCTCTGAG ATGGTGTGGG TGCGCACATC CCTAGGGTAG GCAGTTATTG 2767 GGGGGACCCT TTCTATCCCT TGAACCTCTC ACGTAAAGGG ACTTCCCCAG TCCTGGTTGG 2827 CTTTTGGAAC CTGGTCCTTC TTGCTGTTTT TTACCCTTCC CCGTTTCTAT TACTGCGTGT 2887 AACGTAAAGT GTATCTGAGT GAGGGTGGTG GGAACCCTCT GTCCAGTGCT GTCTCTGTCT 2947 CCTATGGCCT GTGAGTTTCC TTCTAGAGTT CTACTCTTCT CCACCTCTTT GCTCATACAG 3007 AGCTGTGGCC TTGGCGCCTG CCCTGCTTCT GCAGTGCTTC ATTGTCTCGT AGCTTGTCAG 3067 CTGACACCGG AACCGCGGAC GGGACGAAGA CGTCACGAAG TAACAGAGCA TCGAACAGTC 3127 GATTTGTATT GATGTATGTG CCAATGTGGG AAATAAAGAC CTTGTGAGAT AAAAAAAAAA 3187 AAAAAAAAAA AAAA 3201 (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 629 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: Met Arg Pro Lys Asp Gin Val Gin Ser Ser Ala Gly Asp Gly Thr Gly 1 5 10 15 Ser Gly Asp Pro Ala Thr Gly Thr Pro Thr Thr Gin Pro Ala Ala Asp 20 25 30 Pro Ala Pro Glu Pro Ser Ala Glu Pro Lys Pro Ala Pro Ala Gin Gly 35 40 45 Thr Gly Ser Gly Gin Lys Ser Gly Ser Arg Thr Lys Thr Gly Gly Ser 50 55 60 Phe Cys Arg Ser Arg Ile Arg Gly Asp Ser Asp Ala Pro Trp Thr Arg 65 70 75 80 Tyr Ile Phe Gin Gly Pro Tyr Gly Pro Arg Ala Thr Gly Leu Gly Thr 85 90 95 Gly Arg Ala Glu Gly Ile Trp Lys Thr Pro Ala Ala Tyr Ile Gly Arg 100 105 110 Arg Pro Gly Val Ser Gly Pro Glu Arg Ala Ala Phe Ile Arg Glu Leu 115 120 125 Gin Glu Ala Leu Cys Pro Asn Pro Leu Pro Arg Lys Lys Ile Thr Glu 130 135 140 Aβp Aβp Ile Lys Val Met Leu Tyr Leu Leu Glu Glu Lys Glu Arg Asp 145 150 155 160 Leu Aβn Thr Ala Ala Arg Ile Gly Gin Ser Leu Val Lys Gin Aβn Ser 165 170 175 Val Leu Met Glu Glu Asn Asn Lys Leu Glu Thr Met Leu Gly Ser Ala 180 185 190 Arg Glu Glu Ile Leu His Leu Arg Lys Gin Val Asn Leu Arg Asp Asp 195 200 205 Leu Leu Gin Leu Tyr Ser Asp Ser Asp Asp Asp Glu Glu Asp Glu Glu 210 215 220 Aβp Glu Glu Glu Glu Glu Gly Glu Glu Glu Glu Arg Glu Gly Gin Arg 225 230 235 240 Aβp Gin Aβp Gin Gin His Asp His Pro Tyr Gly Ala Pro Lys Pro Pro 245 250 255 Pro Lys Ala Glu Thr Leu His His Cys Pro Gin Leu Glu Ala Leu Lys 260 265 270 Gin Lys Leu Lye Leu Leu Glu Glu Glu Aβn Aβp His Leu Arg Glu Glu 275 280 285 Ala Ser His Leu Asp Asn Leu Glu Asp Lys Glu Gin Met Leu Ile Leu 290 295 300 Glu Cys Val Glu Gin Phe Ser Glu Ala Ser Gin Gin Met Ala Glu Leu 305 310 315 320 Ser Glu Val Leu Val Leu Arg Leu Glu Gly Tyr Glu Arg Gin Gin Lys 325 330 335 Glu Ile Thr Gin Leu Gin Ala Glu Ile Thr Lys Leu Gin Gin Arg Cys 340 345 350 Gin Ser Tyr Gly Ala Gin Thr Glu Lys Leu Gin Gin Gin Leu Ala Ser 355 360 365 Glu Lys Gly Val His Pro Glu Ser Leu Arg Ala Gly Ser His Met Gin 370 375 380 Aβp Tyr Gly Ser Arg Pro Arg Glu Arg Gin Glu Asp Gly Lye Ser Hie 385 390 395 400 Arg Gin Arg Ser Ser Met Pro Ala Gly Ser Val Thr His Tyr Gly Tyr 405 410 415 Ser Val Pro Leu Asp Ala Leu Pro Ser Phe Pro Glu Thr Leu Ala Glu 420 425 430 Glu Leu Arg Thr Ser Leu Arg Lys Phe Ile Thr Asp Pro Ala Tyr Phe 435 440 445 Met Glu Arg Cys Asp Thr Arg Cys Arg Glu Glu Arg Lys Lys Glu Gin 450 455 460 Gly Thr Met Pro Pro Pro Pro Val Gin Asp Leu Lys Pro Pro Glu Asp 465 470 475 480 Phe Glu Ala Pro Glu Glu Leu Val Pro Glu Glu Glu Leu Gly Ala Ile 485 490 495 Glu Glu Val Gly Thr Ala Glu Asp Gly Pro Ala Glu Glu Thr Glu Gin 500 505 510 Ala Ser Glu Glu Thr Glu Ala Trp Glu Glu Val Glu Pro Glu Val Asp 515 520 525 Glu Ala Thr Arg Met Asn Val Val Val Ser Ala Leu Glu Ala Ser Gly 530 535 540 Leu Gly Pro Ser His Leu Asp Met Lys Tyr Val Leu Gin Gin Leu Ser 545 550 555 560 Asn Trp Gin Asp Ala His Ser Lys Arg Gin Gin Lys Gin Lys Val Val 565 570 575 Pro Lys Aβp Ser Pro Ala Pro Gin Gin Gin Thr Asn Met Gly Gly Gly 580 585 590 Ile Val Glu Gin Gin Pro Ile Val Pro Thr Gin Asp Ser Gin Arg Leu 595 600 605 Glu Glu Asp Arg Ala Thr His Ser Pro Ser Ala Arg Glu Glu Glu Gly 610 615 620 Pro Ser Gly Ala Thr 625 (2) INFORMATION FOR SEQ ID NO:7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 378 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..378 (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: GAG GAG GAG AAT CAT CAG CTG AGA GAA GAG GCC TCT CAA CTC GAC ACT 48 Glu Glu Glu Asn His Gin Leu Arg Glu Glu Ala Ser Gin Leu Asp Thr 1 5 10 15 CTT GAG GAT GAG GAA CAG ATG CTC ATT CTG GAG TGT GTG GAG CAG TTT 96 Leu Glu Asp Glu Glu Gin Met Leu Ile Leu Glu Cys Val Glu Gin Phe 20 25 30 TCT GAA GCC AGC CAG CAG ATG GCA GAG CTA TCC GAG GTG TTG GTG CTG 144 Ser Glu Ala Ser Gin Gin Met Ala Glu Leu Ser Glu Val Leu Val Leu 35 40 45 AGG CTG GAA GGC TAT GAG AGG CAG CAG AAG GAG ATC ACT CAG CTG CAG 192 Arg Leu Glu Gly Tyr Glu Arg Gin Gin Lys Glu Ile Thr Gin Leu Gin 50 55 60 GCC GAG ATC ACC AAG CTA CAA CAG CGT TGT CAG TCT TAT GGG GCC CAG 240 Ala Glu Ile Thr Lys Leu Gin Gin Arg Cys Gin Ser Tyr Gly Ala Gin 65 70 75 80 ACG GAG AAA CTG CAG CAG CAG CTG GCC TCA GAG AAG GGA GTC CAC CCA 288 Thr Glu Lys Leu Gin Gin Gin Leu Ala Ser Glu Lys Gly Val His Pro 85 90 95 GAG AGC CTG CGA GCT GGC TCC CAC ATG CAG GAT TAT GGA AGC AGG CCT 336 Glu Ser Leu Arg Ala Gly Ser His Met Gin Aβp Tyr Gly Ser Arg Pro 100 105 110 CGT GAA CGC CAG GAG GAT GGG AAG AGC CAT CGT CAG CGT TCC 378 Arg Glu Arg Gin Glu Aβp Gly Lye Ser His Arg Gin Arg Ser 115 120 125 (2) INFORMATION FOR SEQ ID NO:8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 126 amino acids (B) TYPE: amino acid (0) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: Glu Glu Glu Asn His Gin Leu Arg Glu Glu Ala Ser Gin Leu Asp Thr 1 5 10 15 Leu Glu Asp Glu Glu Gin Met Leu Ile Leu Glu Cys Val Glu Gin Phe 20 25 30 Ser Glu Ala Ser Gin Gin Met Ala Glu Leu Ser Glu Val Leu Val Leu 35 40 45 Arg Leu Glu Gly Tyr Glu Arg Gin Gin Lys Glu Ile Thr Gin Leu Gin 50 55 60 Ala Glu Ile Thr Lys Leu Gin Gin Arg Cys Gin Ser Tyr Gly Ala Gin 65 70 75 80 Thr Glu Lys Leu Gin Gin Gin Leu Ala Ser Glu Lys Gly Val His Pro 85 90 95 Glu Ser Leu Arg Ala Gly Ser His Met Gin Asp Tyr Gly Ser Arg Pro 100 105 110 Arg Glu Arg Gin Glu Asp Gly Lys Ser His Arg Gin Arg Ser 115 120 125 (2) INFORMATION FOR SEQ ID NO:9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10348 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Homo sapiens (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: TTGCTGTGTG AGGCAGAACC TGCGGGGGCA GGGGCGGGCT GGTTCCCTGG CCAGCCATTG 60 GCAGAGTCCG CAGGCTAGGG CTGTCAATCA TGCTGGCCGG CGTGGCCCCG CCTCCGCCGG 120 CGCGGCCCCG CCTCCGCCGG CGCACGTCTG GGACGCAAGG CGCCGTGGGG GCTGCCGGGA 180 CGGGTCCAAG ATGGACGGCC GCTCAGGTTC TGCTTTTACC TGCGGCCCAG AGCCCCATTC 240 ATTGCCCCGG TGCTGAGCGG CGCCGCGAGT CGGCCCGAGG CCTCCGGGGA CTGCCGTGCC 300 GGGCGGGAGA CCGCCATGGC GACCCTGGAA AAGCTGATGA AGGCCTTCGA GTCCCTCAAG 360 TCCTTCCAGC AGCAGCAGCA GCAGCAGCAG CAGCAGCAGC AGCAGCAGCA GCAGCAGCAG 420 CAGCAGCAGC AACAGCCGCC ACCGCCGCCG CCGCCGCCGC CGCCTCCTCA GCTTCCTCAG 480 CCGCCGCCGC AGGCACAGCC GCTGCTGCCT CAGCCGCAGC CGCCCCCGCC GCCGCCCCCG 540 CCGCCACCCG GCCCGGCTGT GGCTGAGGAG CCGCTGCACC GACCAAAGAA AGAACTTTCA 600 GCTACCAAGA AAGACCGTGT GAATCATTGT CTGACAATAT GTGAAAACAT AGTGGCACAG 660 TCTGTCAGAA ATTCTCCAGA ATTTCAGAAA CTTCTGGGCA TCGCTATGGA ACTTTTTCTG 720 CTGTGCAGTG ATGACGCAGA GTCAGATGTC AGGATGGTGG CTGACGAATG CCTCAACAAA 780 GTTATCAAAG CTTTGATGGA TTCTAATCTT CCAAGGTTAC AGCTCGAGCT CTATAAGGAA 840 ATTAAAAAGA ATGGTGCCCC TCGGAGTTTG CGTGCTGCCC TGTGGAGGTT TGCTGAGCTG 900 GCTCACCTGG TTCGGCCTCA GAAATGCAGG CCTTACCTGG TGAACCTTCT GCCGTGCCTG 960 ACTCGAACAA GCAAGAGACC CGAAGAATCA GTCCAGGAGA CCTTGGCTGC AGCTGTTCCC 1020 AAAATTATGG CTTCTTTTGG CAATTTTGCA AATGACAATG AAATTAAGGT TTTGTTAAAG 1080 GCCTTCATAG CGAACCTGAA GTCAAGCTCC CCCACCATTC GGCGGACAGC GGCTGGATCA 1140 GCAGTGAGCA TCTGCCAGCA CTCAAGAAGG ACACAATATT TCTATAGTTG GCTACTAAAT 1200 GTGCTCTTAG GCTTACTCGT TCCTGTCGAG GATGAACACT CCACTCTGCT GATTCTTGGC 1260 GTGCTGCTCA CCCTGAGGTA TTTGGTGCCC TTGCTGCAGC AGCAGGTCAA GGACACAAGC 1320 CTGAAAGGCA GCTTCGGAGT GACAAGGAAA GAAATGGAAG TCTCTCCTTC TGCAGAGCAG 1380 CTTGTCCAGG TTTATGAACT GACGTTACAT CATACACAGC ACCAAGACCA CAATGTTGTG 1440 ACCGGAGCCC TGGAGCTGTT GCAGCAGCTC TTCAGAACGC CTCCACCCGA GCTTCTGCAA 1500 ACCCTGACCG CAGTCGGGGG CATTGGGCAG CTCACCGCTG CTAAGGAGGA GTCTGGTGGC 1560 CGAAGCCGTA GTGGGAGTAT TGTGGAACTT ATAGCTGGAG GGGGTTCCTC ATGCAGCCCT 1620 GTCCTTTCAA GAAAACAAAA AGGCAAAGTG CTCTTAGGAG AAGAAGAAGC CTTGGAGGAT 1680 GACTCTGAAT CGAGATCGGA TGTCAGCAGC TCTGCCTTAA CAGCCTCAGT GAAGGATGAG 1740 ATCAGTGGAG AGCTGGCTGC TTCTTCAGGG GTTTCCACTC CAGGGTCAGC AGGTCATGAC 1800 ATCATCACAG AACAGCCACG GTCACAGCAC ACACTGCAGG CGGACTCAGT GGATCTGGCC 1860 AGCTGTGACT TGACAAGCTC TGCCACTGAT GGGGATGAGG AGGATATCTT GAGCCACAGC 1920 TCCAGCCAGG TCAGCGCCGT CCCATCTGAC CCTGCCATGG ACCTGAATGA TGGGACCCAG 1980 GCCTCGTCGC CCATCAGCGA CAGCTCCCAG ACCACCACCG AAGGGCCTGA TTCAGCTGTT 2040 ACCCCTTCAG ACAGTTCTGA AATTGTGTTA GACGGTACCG ACAACCAGTA TTTGGGCCTG 2100 CAGATTGGAC AGCCCCAGGA TGAAGATGAG GAAGCCACAG GTATTCTTCC TGATGAAGCC 2160 TCGGAGGCCT TCAGGAACTC TTCCATGGCC CTTCAACAGG CACATTTATT GAAAAACATG 2220 AGTCACTGCA GGCAGCCTTC TGACAGCAGT GTTGATAAAT TTGTGTTGAG AGATGAAGCT 2280 ACTGAACCGG GTGATCAAGA AAACAAGCCT TGCCGCATCA AAGGTGACAT TGGACAGTCC 2340 ACTGATGATG ACTCTGCACC TCTTGTCCAT TGTGTCCGCC TTTTATCTGC TTCGTTTTTG 2400 CTAACAGGGG GAAAAAATGT GCTGGTTCCG GACAGGGATG TGAGGGTCAG CGTGAAGGCC 2460 CTGGCCCTCA GCTGTGTGGG AGCAGCTGTG GCCCTCCACC CGGAATCTTT CTTCAGCAAA 2520 CTCTATAAAG TTCCTCTTGA CACCACGGAA TACCCTGAGG AACAGTATGT CTCAGACATC 2580 TTGAACTACA TCGATCATGG AGACCCACAG GTTCGAGGAG CCACTGCCAT TCTCTGTGGG 2640 ACCCTCATCT GCTCCATCCT CAGCAGGTCC CGCTTCCACG TGGGAGATTG GATGGGCACC 2700 ATTAGAACCC TCACAGGAAA TACATTTTCT TTGGCGGATT GCATTCCTTT GCTGCGGAAA 2760 ACACTGAAGG ATGAGTCTTC TGTTACTTGC AAGTTAGCTT GTACAGCTGT GAGGAACTGT 2820 GTCATGAGTC TCTGCAGCAG CAGCTACAGT GAGTTAGGAC TGCAGCTGAT CATCGATGTG 2880 CTGACTCTGA GGAACAGTTC CTATTGGCTG GTGAGGACAG AGCTTCTGGA AACCCTTGCA 2940 GAGATTGACT TCAGGCTGGT GAGCTTTTTG GAGGCAAAAG CAGAAAACTT ACACAGAGGG 3000 GCTCATCATT ATACAGGGCT TTTAAAACTG CAAGAACGAG TGCTCAATAA TGTTGTCATC 3060 CATTTGCTTG GAGATGAAGA CCCCAGGGTG CGACATGTTG CCGCAGCATC ACTAATTAGG 3120 CTTGTCCCAA AGCTGTTTTA TAAATGTGAC CAAGGACAAG CTGATCCAGT AGTGGCCGTG 3180 GCAAGAGATC AAAGCAGTGT TTACCTGAAA CTTCTCATGC ATGAGACGCA GCCTCCATCT 3240 CATTTCTCCG TCAGCACAAT AACCAGAATA TATAGAGGCT ATAACCTACT ACCAAGCATA 3300 ACAGACGTCA CTATGGAAAA TAACCTTTCA AGAGTTATTG CAGCAGTTTC TCATGAACTA 3360 ATCACATCAA CCACCAGAGC ACTCACATTT GGATGCTGTG AAGCTTTGTG TCTTCTTTCC 3420 ACTGCCTTCC CAGTTTGCAT TTGGAGTTTA GGTTGGCACT GTGGAGTGCC TCCACTGAGT 3480 GCCTCAGATG AGTCTAGGAA GAGCTGTACC GTTGGGATGG CCACAATGAT TCTGACCCTG 3540 CTCTCGTCAG CTTGGTTCCC ATTGGATCTC TCAGCCCATC AAGATGCTTT GATTTTGGCC 3600 GGAAACTTGC TTGCAGCCAG TGCTCCCAAA TCTCTGAGAA GTTCATGGGC CTCTGAAGAA 3660 GAAGCCAACC CAGCAGCCAC CAAGCAAGAG GAGGTCTGGC CAGCCCTGGG GGACCGGGCC 3720 CTGGTGCCCA TGGTGGAGCA GCTCTTCTCT CACCTGCTGA AGGTGATTAA CATTTGTGCC 3780 CACGTCCTGG ATGACGTGGC TCCTGGACCC GCAATAAAGG CAGCCTTGCC TTCTCTAACA 3840 AACCCCCCTT CTCTAAGTCC CATCCGACGA AAGGGGAAGG AGAAAGAACC AGGAGAACAA 3900 GCATCTGTAC CGTTGAGTCC CAAGAAAGGC AGTGAGGCCA GTGCAGCTTC TAGACAATCT 3960 GATACCTCAG GTCCTGTTAC AACAAGTAAA TCCTCATCAC TGGGGAGTTT CTATCATCTT 4020 CCTTCATACC TCAAACTGCA TGATGTCCTG AAAGCTACAC ACGCTAACTA CAAGGTCACG 4080 CTGGATCTTC AGAACAGCAC GGAAAAGTTT GGAGGGTTTC TCCGCTCAGC CTTGGATGTT 4140 CTTTCTCAGA TACTAGAGCT GGCCACACTG CAGGACATTG GGAAGTGTGT TGAAGAGATC 4200 CTAGGATACC TGAAATCCTG CTTTAGTCGA GAACCAATGA TGGCAACTGT TTGTGTTCAA 4260 CAATTGTTGA AGACTCTCTT TGGCACAAAC TTGGCCTCCC AGTTTGATGG CTTATCTTCC 4320 AACCCCAGCA AGTCACAAGG CCGAGCACAG CGCCTTGGCT CCTCCAGTGT GAGGCCAGGC 4380 TTGTACCACT ACTGCTTCAT GGCCCCGTAC ACCCACTTCA CCCAGGCCCT CGCTGACGCC 4440 AGCCTGAGGA ACATGGTGCA GGCGGAGCAG GAGAACGACA CCTCGGGATG GTTTGATGTC 4500 CTCCAGAAAG TGTCTACCCA GTTGAAGACA AACCTCACGA GTGTCACAAA GAACCGTGCA 4560 GATAAGAATG CTATTCATAA TCACATTCGT TTGTTTGAAC CTCTTGTTAT AAAAGCTTTA 4620 AAACAGTACA CGACTACAAC ATGTGTGCAG TTACAGAAGC AGGTTTTAGA TTTGCTGGCG 4680 CAGCTGGTTC AGTTACGGGT TAATTACTGT CTTCTGGATT CAGATCAGGT GTTTATTGGC 4740 TTTGTATTGA AACAGTTTGA ATACATTGAA GTGGGCCAGT TCAGGGAATC AGAGGCAATC 4800 ATTCCAAACA TCTTTTTCTT CTTGGTATTA CTATCTTATG AACGCTATCA TTCAAAACAG 4860 ATCATTGGAA TTCCTAAAAT CATTCAGCTC TGTGATGGCA TCATGGCCAG TGGAAGGAAG 4920 GCTGTGACAC ATGCCATACC GGCTCTGCAG CCCATAGTCC ACGACCTCTT TGTATTAAGA 4980 GGAACAAATA AAGCTGATGC AGGAAAAGAG CTTGAAACCC AAAAAGAGGT GGTGGTGTCA 5040 ATGTTACTGA GACTCATCCA GTACCATCAG GTGTTGGAGA TGTTCATTCT TGTCCTGCAG 5100 CAGTGCCACA AGGAGAATGA AGACAAGTGG AAGCGACTGT CTCGACAGAT AGCTGACATC 5160 ATCCTCCCAA TGTTAGCCAA ACAGCAGATG CACATTGACT CTCATGAAGC CCTTGGAGTG 5220 TTAAATACAT TATTTGAGAT TTTGGCCCCT TCCTCCCTCC GTCCGGTAGA CATGCTTTTA 5280 CGGAGTATGT TCGTCACTCC AAACACAATG GCGTCCGTGA GCACTGTTCA ACTGTGGATA 5340 TCGGGAATTC TGGCCATTTT GAGGGTTCTG ATTTCCCAGT CAACTGAAGA TATTGTTCTT 5400 TCTCGTATTC AGGAGCTCTC CTTCTCTCCG TATTTAATCT CCTGTACAGT AATTAATAGG 5460 TTAAGAGATG GGGACAGTAC TTCAACGCTA GAAGAACACA GTGAAGGGAA ACAAATAAAG 5520 AATTTGCCAG AAGAAACATT TTCAAGGTTT CTATTACAAC TGGTTGGTAT TCTTTTAGAA 5580 GACATTGTTA CAAAACAGCT GAAGGTGGAA ATGAGTGAGC AGCAACATAC TTTCTATTGC 5640 CAGGAACTAG GCACACTGCT AATGTGTCTG ATCCACATCT TCAAGTCTGG AATGTTCCGG 5700 AGAATCACAG CAGCTGCCAC TAGGCTGTTC CGCAGTGATG GCTGTGGCGG CAGTTTCTAC 5760 ACCCTGGACA GCTTGAACTT GCGGGCTCGT TCCATGATCA CCACCCACCC GGCCCTGGTG 5820 CTGCTCTGGT GTCAGATACT GCTGCTTGTC AACCACACCG ACTACCGCTG GTGGGCAGAA 5880 GTGCAGCAGA CCCCGAAAAG ACACAGTCTG TCCAGCACAA AGTTACTTAG TCCCCAGATG 5940 TCTGGAGAAG AGGAGGATTC TGACTTGGCA GCCAAACTTG GAATGTGCAA TAGAGAAATA 6000 GTACGAAGAG GGGCTCTCAT TCTCTTCTGT GATTATGTCT GTCAGAACCT CCATGACTCC 6060 GAGCACTTAA CGTGGCTCAT TGTAAATCAC ATTCAAGATC TGATCAGCCT TTCCCACGAG 6120 CCTCCAGTAC AGGACTTCAT CAGTGCCGTT CATCGGAACT CTGCTGCCAG CGGCCTGTTC 6180 ATCCAGGCAA TTCAGTCTCG TTGTGAAAAC CTTTCAACTC CAACCATGCT GAAGAAAACT 6240 CTTCAGTGCT TGGAGGGGAT CCATCTCAGC CAGTCGGGAG CTGTGCTCAC GCTGTATGTG 6300 GACAGGCTTC TGTGCACCCC TTTCCGTGTG CTGGCTCGCA TGGTCGACAT CCTTGCTTGT 6360 CGCCGGGTAG AAATGCTTCT GGCTGCAAAT TTACAGAGCA GCATGGCCCA GTTGCCAATG 6420 GAAGAACTCA ACAGAATCCA GGAATACCTT CAGAGCAGCG GGCTCGCTCA GAGACACCAA 6480 AGGCTCTATT CCCTGCTGGA CAGGTTTCGT CTCTCCACCA TGCAAGACTC ACTTAGTCCC 6540 TCTCCTCCAG TCTCTTCCCA CCCGCTGGAC GGGGATGGGC ACGTGTCACT GGAAACAGTG 6600 AGTCCGGACA AAGACTGGTA CGTTCATCTT GTCAAATCCC AGTGTTGGAC CAGGTCAGAT 6660 TCTGCACTGC TGGAAGGTGC AGAGCTGGTG AATCGGATTC CTGCTGAAGA TATGAATGCC 6720 TTCATGATGA ACTCGGAGTT CAACCTAAGC CTGCTAGCTC CATGCTTAAG CCTAGGGATG 6780 AGTGAAATTT CTGGTGGCCA GAAGAGTGCC CTTTTTGAAG CAGCCCGTGA GGTGACTCTG 6840 GCCCGTGTGA GCGGCACCGT GCAGCAGCTC CCTGCTGTCC ATCATGTCTT CCAGCCCGAG 6900 CTGCCTGCAG AGCCGGCGGC CTACTGGAGC AAGTTGAATG ATCTGTTTGG GGATGCTGCA 6960 CTGTATCAGT CCCTGCCCAC TCTGGCCCGG GCCCTGGCAC AGTACCTGGT GGTGGTCTCC 7020 AAACTGCCCA GTCATTTGCA CCTTCCTCCT GAGAAAGAGA AGGACATTGT GAAATTCGTG 7080 GTGGCAACCC TTGAGGCCCT GTCCTGGCAT TTGATCCATG AGCAGATCCC GCTGAGTCTG 7140 GATCTCCAGG CAGGGCTGGA CTGCTGCTGC CTGGCCCTGC AGCTGCCTGG CCTCTGGAGC 7200 GTGGTCTCCT CCACAGAGTT TGTGACCCAC GCCTGCTCCC TCATCTACTG TGTGCACTTC 7260 ATCCTGGAGG CCGTTGCAGT GCAGCCTGGA GAGCAGCTTC TTAGTCCAGA AAGAAGGACA 7320 AATACCCCAA AAGCCATCAG CGAGGAGGAG GAGGAAGTAG ATCCAAACAC ACAGAATCCT 7380 AAGTATATCA CTGCAGCCTG TGAGATGGTG GCAGAAATGG TGGAGTCTCT GCAGTCGGTG 7440 TTGGCCTTGG GTCATAAAAG GAATAGCGGC GTGCCGGCGT TTCTCACGCC ATTGCTCAGG 7500 AACATCATCA TCAGCCTGGC CCGCCTGCCC CTTGTCAACA GCTACACACG TGTGCCCCCA 7560 CTGGTGTGGA AGCTTGGATG GTCACCCAAA CCGGGAGGGG ATTTTGGCAC AGCATTCCCT 7620 GAGATCCCCG TGGAGTTCCT CCAGGAAAAG GAAGTCTTTA AGGAGTTCAT CTACCGCATC 7680 AACACACTAG GCTGGACCAG TCGTACTCAG TTTGAAGAAA CTTGGGCCAC CCTCCTTGGT 7740 GTCCTGGTGA CGCAGCCCCT CGTGATGGAG CAGGAGGAGA GCCCACCAGA AGAAGACACA 7800 GAGAGGACCC AGATCAACGT CCTGGCCGTG CAGGCCATCA CCTCACTGGT GCTCAGTGCA 7860 ATGACTGTGC CTGTGGCCGG CAACCCAGCT GTAAGCTGCT TGGAGCAGCA GCCCCGGAAC 7920 AAGCCTCTGA AAGCTCTCGA CACCAGGTTT GGGAGGAAGC TGAGCATTAT CAGAGGGATT 7980 GTGGAGCAAG AGATTCAAGC AATGGTTTCA AAGAGAGAGA ATATTGCCAC CCATCATTTA 8040 TATCAGGCAT GGGATCCTGT CCCTTCTCTG TCTCCGGCTA CTACAGGTGC CCTCATCAGC 8100 CACGAGAAGC TGCTGCTACA GATCAACCCC GAGCGGGAGC TGGGGAGCAT GAGCTACAAA 8160 CTCGGCCAGG TGTCCATACA CTCCGTGTGG CTGGGGAACA GCATCACACC CCTGAGGGAG 8220 GAGGAATGGG ACGAGGAAGA GGAGGAGGAG GCCGACGCCC CTGCACCTTC GTCACCACCC 8280 ACGTCTCCAG TCAACTCCAG GAAACACCGG GCTGGAGTTG ACATCCACTC CTGTTCGCAG 8340 TTTTTGCTTG AGTTGTACAG CCGCTGGATC CTGCCGTCCA GCTCAGCCAG GAGGACCCCG 8400 GCCATCCTGA TCAGTGAGGT GGTCAGATCC CTTCTAGTGG TCTCAGACTT GTTCACCGAG 8460 CGCAACCAGT TTGAGCTGAT GTATGTGACG CTGACAGAAC TGCGAAGGGT GCACCCTTCA 8520 GAAGACGAGA TCCTCGCTCA GTACCTGGTG CCTGCCACCT GCAAGGCAGC TGCCGTCCTT 8580 GGGATGGACA AGGCCGTGGC GGAGCCTGTC AGCCGCCTGC TGGAGAGCAC GCTCAGGAGC 8640 AGCCACCTGC CCAGCAGGGT TGGAGCCCTG CACGGCGTCC TCTATGTGCT GGAGTGCGAC 8700 CTGCTGGACG ACACTGCCAA GCAGCTCATC CCGGTCATCA GCGACTATCT CCTCTCCAAC 8760 CTGAAAGGGA TCGCCCACTG CGTGAACATT CACAGCCAGC AGCACGTACT GGTCATGTGT 8820 GCCACTGCGT TTTACCTCAT TGAGAACTAT CCTCTGGACG TAGGGCCGGA ATTTTCAGCA 8880 TCAATAATAC AGATGTGTGG GGTGATGCTG TCTGGAAGTG AGGAGTCCAC CCCCTCCATC 8940 ATTTACCACT GTGCCCTCAG AGGCCTGGAG CGCCTCCTGC TCTCTGAGCA GCTCTCCCGC 9000 CTGGATGCAG AATCGCTGGT CAAGCTGAGT GTGGACAGAG TGAACGTGCA CAGCCCGCAC 9060 CGGGCCATGG CGGCTCTGGG CCTGATGCTC ACCTGCATGT ACACAGGAAA GGAGAAAGTC 9120 AGTCCGGGTA GAACTTCAGA CCCTAATCCT GCAGCCCCCG ACAGCGAGTC AGTGATTGTT 9180 GCTATGGAGC GGGTATCTGT TCTTTTTGAT AGGATCAGGA AAGGCTTTCC TTGTGAAGCC 9240 AGAGTGGTGG CCAGGATCCT GCCCCAGTTT CTAGACGACT TCTTCCCACC CCAGGACATC 9300 ATGAACAAAG TCATCGGAGA GTTTCTGTCC AACCAGCAGC CATACCCCCA GTTCATGGCC 9360 ACCGTGGTGT ATAAGGTGTT TCAGACTCTG CACAGCACCG GGCAGTCGTC CATGGTCCGG 9420 GACTGGGTCA TGCTGTCCCT CTCCAACTTC ACGCAGAGGG CCCCGGTCGC CATGGCCACG 9480 TGGAGCCTCT CCTGCTTCTT TGTCAGCGCG TCCACCAGCC CGTGGGTCGC GGCGATCCTC 9540 CCACATGTCA TCAGCAGGAT GGGCAAGCTG GAGCAGGTGG ACGTGAACCT TTTCTGCCTG 9600 GTCGCCACAG ACTTCTACAG ACACCAGATA GAGGAGGAGC TCGACCGCAG GGCCTTCCAG 9660 TCTGTGCTTG AGGTGGTTGC AGCCCCAGGA AGCCCATATC ACCGGCTGCT GACTTGTTTA 9720 CGAAATGTCC ACAAGGTCAC CACCTGCTGA GCGCCATGGT GGGAGAGACT GTGAGGCGGC 9780 AGCTGGGGCC GGAGCCTTTG GAAGTCTGTG CCCTTGTGCC CTGCCTCCAC CGAGCCAGCT 9840 TGGTCCCTAT GGGCTTCCGC ACATGCCGCG GGCGGCCAGG CAACGTGCGT GTCTCTGCCA 9900 TGTGGCAGAA GTGCTCTTTG TGGCAGTGGC CAGGCAGGGA GTGTCTGCAG TCCTGGTGGG 9960 GCTGAGCCTG AGGCCTTCCA GAAAGCAGGA GCAGCTGTGC TGCACCCCAT GTGGGTGACC 10020 AGGTCCTTTC TCCTGATAGT CACCTGCTGG TTGTTGCCAG GTTGCAGCTG CTCTTGCATC 10080 TGGGCCAGAA GTCCTCCCTC CTGCAGGCTG GCTGTTGGCC CCTCTGCTGT CCTGCAGTAG 10140 AAGGTGCCGT GAGCAGGCTT TGGGAACACT GGCCTGGGTC TCCCTGGTGG GGTGTGCATG 10200 CCACGCCCCG TGTCTGGATG CACAGATGCC ATGGCCTGTG CTGGGCCAGT GGCTGGGGGT 10260 GCTAGACACC CGGCACCATT CTCCCTTCTC TCTTTTCTTC TCAGGATTTA AAATTTAATT 10320 ATATCAGTAA AGAGATTAAT TTTAACGT 10348

Claims: CLAIMS 1. A method for identifying compounds which interfere with the binding of huntingtin associated protein- 1 (HAPl) to huntingtin, said compounds being candidate therapeutic agents, said method comprising the steps of: contacting: a first protein; a second protein; and a compound to be tested for its capacity to interfere with binding of said first and second proteins to each other; wherein the first protein comprises huntingtin associated protein- 1 (HAPl) and the second protein comprises huntingtin or the first protein comprises huntingtin and the second protein comprises huntingtin associated protein- 1 (HAPl); and determining the quantity of the first protein which is bound to, is displaced from, or is prevented from binding to, the second protein, wherein a compound which diminishes the quantity of the first protein bound to the second protein, or which displaces first protein bound to the second protein, or which prevents first protein from binding to the second protein, is identified as a candidate therapeutic agent. 2. The method of claim 1 wherein an antibody is used to determine the quantity of the first protein which is bound to, is displaced from, or is prevented from binding to, the second protein. 3. The method of claim 1 wherein one of said two proteins is fixed to a solid support. 4. The method of claim 1 wherein one of said two proteins is labeled. 5. The method of claim 1 wherein an antibody specifically immunoreactive with said second protein is used to separate bound first protein from unbound first protein. 6. The method of claim 1 wherein at least one of said first and said second proteins is a fusion protein. 7. The method of claim 1 wherein said first or said second protein consists of huntingtin associated protein- 1 (HAPl) as shown in SEQ ID NO: 8. 8. The method of claim 1 wherein said first or said second protein consists of huntingtin. 9. The method of claim 1 wherein the huntingtin associated protein- 1 (HAPl) is human huntingtin associated protein- 1 (HAPl). 10. The method of claim 1 wherein the huntingtin is human huntingtin. 11. A method for identifying compounds which interfere with the binding of huntingtin associated protein-1 (HAPl) to huntingtin, said compounds being candidate therapeutic agents, said method comprising the steps of: contacting: a first polypeptide; a second polypeptide; and a compound to be tested for its capacity to interfere with binding of said first and second polypeptides to each other; wherein at least one of said first and said second polypeptides is a polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin or huntingtin associated protein- 1 (HAPl) proteins, wherein each of said polypeptides contain a sufficient portion of huntingtin or huntingtin associated protein- 1 (HAPl) proteins to bind to the other polypeptide; wherein when said first polypeptide is huntingtin or a polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin, then said second polypeptide is huntingtin associated protein- 1 (HAPl) or a polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin associated protein- 1 (HAPl); and when said first polypeptide is huntingtin associated protein- 1 (HAPl) or a polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin associated protein- 1 (HAPl), then said second polypeptide is huntingtin or a polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin; and determining the quantity of the first polypeptide which is bound to, is displaced from, or is prevented from binding to, the second polypeptide, wherein a compound which diminishes the quantity of the first protein, or which prevents first protein from binding to the second protein, is identified as a candidate therapeutic agent. 12. The method of claim 11 wherein an antibody is used to determine the quantity of the first polypeptide which is bound to, is displaced from, or is prevented from binding to, the second polypeptide. 13. The method of claim 11 wherein one of said two polypeptides is fixed to a solid support. 14. The method of claim 1 1 wherein an antibody specifically immunoreactive with said second polypeptide is used to separate bound first polypeptide from unbound first polypeptide. 15. The method of claim 11 wherein one of said two polypeptides is labeled. 16. The method of claim 11 wherein said polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin associated protein- 1 (HAPl) comprises amino acids 105-136 of human huntingtin associated protein- 1 (HAPl) as shown in SEQ ID NO: 8. 17. The method of claim 11 wherein said polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin associated protein-1 (HAPl) comprises amino acids 4-136 of human huntingtin associated protein- 1 (HAPl) as shown in SEQ ID NO: 8. 18. The method of claim 11 wherein said polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin associated protein-1 (HAPl) comprises amino acids 50-136 of human huntingtin associated protein- 1 (HAPl) as shown in SEQ ID NO: 8. 19. The method of claim 11 wherein said polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin associated protein- 1 (HAPl) comprises amino acids 75-136 of human huntingtin associated protein-1 (HAPl) as shown in SEQ ID NO:8. 20. The method of claim 11 wherein said polypeptide which comprises less than all of the complete sequence of amino acids of huntingtin comprises amino acids 1-230 of huntingtin as shown in SEQ ID NO: l. 21. The method of claim 11 wherein the huntingtin associated protein- 1 (HAPl) is human huntingtin associated protein- 1 (HAPl). 22. The method of claim 11 wherein the huntingtin is human huntingtin. 23. The method of claim 11 wherein at least one of said first and said second polypeptides is a fusion polypeptide. 24. The method of claim 23 wherein said fusion polypeptide comprises amino acids 105-136 of human huntingtin associated protein- 1 (HAPl) as shown in SEQ ID NO: 8. 25. The method of claim 23 wherein said fusion polypeptide comprises amino acids 4-136 of human huntingtin associated protein- 1 (HAPl) as shown in SEQ ID NO:8. 26. The method of claim 23 wherein said fusion polypeptide comprises amino acids 50-136 of human huntingtin associated protein-1 (HAPl) as shown in SEQ ID NO:8. 27. The method of claim 23 wherein said fusion polypeptide comprises amino acids 75-136 of human huntingtin associated protein- 1 (HAPl) as shown in SEQ ID NO:8. 28. The method of claim 23 wherein said fusion polypeptide comprises amino acids 1-230 of human huntingtin as shown in SEQ ID NO: l. 29. A method of identifying compounds which interfere with the binding of human huntingtin associated protein-1 (HAPl) to human huntingtin, comprising: providing a cell which comprises three recombinant DNA constructs, said first construct encoding a first polypeptide fused to a sequence- specific DNA-binding domain, said second construct encoding a second polypeptide fused to a transcriptional activation domain, said third construct comprising a reporter gene downstream from a DNA element which is recognized by said sequence-specific DNA-binding domain, wherein said first polypeptide is a huntingtin associated protein- 1 (HAPl) polypeptide and said second protein is a huntingtin polypeptide, or said first polypeptide is a huntingtin polypeptide and said second polypeptide is a huntingtin associated protein- 1 (HAPl) polypeptide; wherein said polypeptides contain a sufficient portion of huntingtin and huntingtin associated protein-1 (HAPl) proteins to bind to the other polypeptide; contacting said cell with a compound to be tested for its capacity to inhibit binding of huntingtin to huntingtin associated protein- 1 (HAPl); determining the amount of expression of the reporter gene in the presence of said compound. 30. A cell which comprises three recombinant DNA constructs, said first construct encoding a first polypeptide fused to a sequence-specific DNA- binding domain, said second construct encoding a second polypeptide fused to a transcriptional activation domain, said third construct comprising a reporter gene downstream from a DNA element which is recognized by said sequence-specific DNA-binding domain, wherein said first polypeptide is an huntingtin associated protein- 1 (HAPl) polypeptide and said second protein is a huntingtin polypeptide, or said first polypeptide is a huntingtin polypeptide and said second polypeptide is an huntingtin associated protein- 1 (HAPl) polypeptide; wherein said polypeptides contain a sufficient portion of huntingtin and huntingtin associated protein- 1 (HAPl) proteins to bind to the other polypeptide. 31. A method of determining the quantity of human huntingtin which binds to human huntingtin associated protein- 1 (HAPl), or of human huntingtin associated protein- 1 (HAPl) which binds to human huntingtin, said method comprising: contacting: a first protein and a second protein, wherein the first protein is human huntingtin associated protein- 1 (HAPl) and the second protein is human huntingtin or the first protein is human huntingtin and the second protein is human huntingtin associated protein- 1 (HAPl); and determining the quantity of the first protein which is bound to the second protein. 32. A method of screening for candidate drugs for treating, delaying onset, or preventing Huntington's Disease, comprising the steps of: contacting a compound to be tested with a yeast cell comprising: two fused gene constructs wherein a first construct comprises a yeast GAL-4 binding domain and a domain selected from the group consisting of: a glutamine repeat-containing domain of a huntingtin gene and a huntingtin binding domain of a huntingtin associated protein- 1 (HAPl) gene, and wherein a second construct comprises a yeast GAL-4 activation domain, and a domain selected from the group consisting of: a glutamine repeat-containing domain of a huntingtin gene and a huntingtin binding domain of a huntingtin associated protein- 1 (HAPl) gene, wherein when the first construct comprises a glutamine repeat-containing domain of a huntingtin gene, the second construct comprises a huntingtin binding domain of a huntingtin associated protein- 1 (HAPl) gene, and when the second construct comprises a glutamine repeat-containing domain of a huntingtin gene, the first construct comprises a huntingtin binding domain of a huntingtin associated protein- 1 (HAPl) gene; and a β-galactosidase reporter gene under the control of a yeast GAL-4 promoter, which is activated by the gene products of said two fused gene constructs, and selecting those compounds which diminish the expression of β- galactosidase by said cells. 33. A yeast cell comprising: two fused gene constructs and a β- galactosidase reporter gene under the control of a yeast GAL-4 promoter, wherein a first construct comprises a yeast GAL-4 binding domain and a domain selected from the group consisting of: a glutamine repeat- containing domain of a huntingtin gene and a huntingtin binding domain of a huntingtin associated protein- 1 (HAPl) gene, and a second construct comprises a yeast GAL-4 activation domain, and a domain selected from the group consisting of: a glutamine repeat-containing domain of a huntingtin gene and a huntingtin binding domain of a huntingtin associated protein- 1 (HAPl) gene, wherein when the first construct comprises a glutamine repeat- containing domain of a huntingtin gene, the second construct comprises a huntingtin binding domain of a huntingtin associated protein- 1 (HAPl) gene, and when the second construct comprises a glutamine repeat-containing domain of a huntingtin gene, the first construct comprises a huntingtin binding domain of a huntingtin associated protein- 1 (HAPl) gene; and said /3-galactosidase reporter gene is activated by the gene products of said two fused gene constructs. 34. A cDNA segment encoding at least 12 contiguous amino acids of a mammalian huntingtin associated protein- 1 (HAPl). 35. A cDNA segment consisting of at least 20 contiguous nucleotides of a mammalian gene encoding huntingtin associated protein-1 (HAPl). 36. The cDNA segment of claim 34 wherein said amino acids are selected from consecutive sequences in SEQ ID NO:4. 37. The cDNA segment of claim 35 wherein said nucleotides are selected from consecutive sequences in SEQ ID NO: 3. 38. The cDNA segment of claim 35 wherein said nucleotides are selected from consecutive sequences in SEQ ID NO:5. 39. The cDNA segment of claim 35 wherein said nucleotides are selected from consecutive sequences in SEQ ID NO: 7. 40. The cDNA segment of claim 35 consisting of a sequence as shown in SEQ ID NO:3. 41. The cDNA segment of claim 35 consisting of a sequence as shown in SEQ ID NO:5. 42. The cDNA segment of claim 35 consisting of a sequence as shown in SEQ ID NO:7. 43. A fusion protein consisting of all or a portion of a first and a second protein, wherein said second protein is a mammalian huntingtin associated protein- 1 (HAPl), and the portion of said mammalian huntingtin associated protein- 1 (HAPl) is sufficient to bind to huntingtin. 44. The fusion protein of claim 43 wherein said first protein is yeast GAL-4 and said wherein said portion of said first protein is selected from the group consisting of the binding domain and the activation domain. 45. The fusion protein of claim 43 wherein said second protein is rat HAP1-A as shown in SEQ ID NO:4. 46. The fusion protein of claim 43 wherein said second protein is rat HAP1-B as shown in SEQ ID NO:6. 47. The fusion protein of claim 43 wherein said portion of said second protein is human hHAPl as shown in SEQ ID NO:8. 48. An isolated mammalian protein which is a huntingtin associated protein- 1 (HAPl). 49. The protein of claim 47 which consists of the sequence shown in SEQ ID NO:4. 50. The protein of claim 47 which consists of the sequence shown in SEQ ID NO:6. 51. The protein of claim 47 which consists of the sequence shown in SEQ ID NO:8. 52. The protein of claim 47 which comprises the sequence shown in SEQ ID NO: 8. 53. A preparation of antibodies which are specifically immunoreactive with a huntingtin associated protein- 1 (HAPl). 54. The preparation of claim 53 wherein the antibodies are polyclonal. 55. The preparation of claim 53 wherein the antibodies are monoclonal.

Inventors: LANAHAN ANTHONY, LI SHI-HUA, LI XIAO-JIANG, ROSS CHRISTOPHER A, SHARP ALAN H, SNYDER SOLOMON H, WORLEY PAUL F, LANAHAN, ANTHONY, LI, SHI-HUA, LI, XIAO-JIANG, ROSS, CHRISTOPHER, A., SHARP, ALAN, H., SNYDER, SOLOMON, H., WORLEY, PAUL, F.
Applicants: UNIV JOHNS HOPKINS, THE JOHNS HOPKINS UNIVERSITY
Application year: 1996
Main classification: C12N15
IPC codes: C07K14, C07K17, C07K19, C12N1, C12N5, C12N15, C12Q1, G01N33