Abstract:
The present invention discloses complexes of cellular signaling proteins that interact in vivo with the HIV-encoded auxiliary proteins Nef and Tat to modulate their activity. This complex includes the novel serine/threonine kinase PAK4 and the novel guanine nucleotide exchange factor Cdc42-GEF, which synergize to stimulate Tat transcriptional activity, and the acetyl-transferase Tip60 which modifies Nef. These cellular partners of the HIV auxiliary proteins represent novel targets for HIV therapeutics. The invention provides isolated DNA and vectors encoding PAK4 and Cdc42-GEF, and methods of producing recombinant forms of these proteins. The invention also provides methods for identifying compounds that modulate the activity of HIV-Tat, HIV-Nef or Tip60, and methods for modulating the activity of these enzymes.

Description:
RELATED APPLICATIONS  
       [0001]    This application is a continuation-in-part of, and claims priority to, U.S. Ser. No. 09/750,457, filed Dec. 28, 2000, presently pending, and U.S. Ser. No. 60/173,939, filed Dec. 30, 1999, now abandoned, the disclosures of which are incorporated by reference herein. 
     
    
     
       FIELD OF THE INVENTION  
         [0002]    The invention relates generally to protein-protein interactions, and more particularly to interactions involving viral transcriptional enzymes.  
         BACKGROUND OF THE INVENTION  
         [0003]    The handful of proteins encoded by the genome of the Human Immunodeficiency Virus (HIV) have been the object of intense scrutiny for clues as to the mechanism of the disease AIDS and as targets for potential AIDS therapeutics. Today&#39;s most effective drugs in the treatment of AIDS were designed either as inhibitors of the HIV-encoded DNA polymerase or the HIV protease. However there are a number of other, smaller, HIV-encoded proteins, so-called auxiliary proteins, whose function still remains unclear (Cullen (1998)  Cell  93:685-692). These proteins and their cellular effector proteins represent additional potential targets for the development of future AIDS therapeutics.  
           [0004]    One of the most poorly understood of these auxiliary proteins is HIV-Nef (reviewed in Marsh (1999)  Arch Biochem Biophys.  365(2):192-198). Among several putative cellular Nef-binding proteins that may hold clues as to its in vivo function is a cellular serine/threonine kinase activity (Nef-associated kinase or NAK, reviewed in Trono and Wang (1997)  Chemistry and Biology  4:13-15). Nef is thought to activate NAK which may help mediate the role Nef plays in HIV replication (Lu et al. (1996)  Current Biology  6(12):1677-1684). NAK is believed to be a member of the PAK or p21-activated kinase family by virtue of its cross-reactivity with anti-PAK antibodies and its regulation by Cdc42 and Rac.  
           [0005]    As presently disclosed, NAK has now been identified as a novel member of the PAK family presently named “PAK4” (this protein has since been independently identified by Abo et al. (1998)  EMBO Journal  17(22):6527-6540). The PAK family of kinases represents a growing group of kinases related in the sequence of their kinase domains and in their binding and regulation by Rho-family small G proteins such as Rac and Cdc42 (reviewed in Manser and Lim (1999)  Progress in Molecular Subcellular Biology  22:115-133; Knaus and Bokoch (1998)  Intl. J. Biochem. Cell Biol.  30:857-862). The three PAK isoforms characterized in mammals to date have been designated Pak1 (also known as alpha-Pak), Pak2 (also known as gamma-Pak), and Pak3 (beta-Pak). Pak1 and Pak3 are expressed primarily in the brain, while Pak2 tissue expression is ubiquitous. These Pak family members have been characterized primarily in terms of their role in regulation of the cytoskeleton and morphogenesis, and as upstream activators of the JNK and p38 MAPK pathways. Recently NAK was reported to be identical to Pak2 (Renkema et al. (1999)  Current Biology  9:1407-1410); however it was not possible to reproduce the interaction between Nef and Pak2 described in that report and instead Pak4 was identified as the Nef-associated kinase.  
           [0006]    Pak1 and Pak3 have previously been shown to bind proteins with guanine nucleotide exchange factor activity, members of the so-called Cool/Pix family (reviewed in Bagrodia and Cerione (1999)  Trends in Cell Biology  9:350-355). These enzymes presumably facilitate Pak activation by charging Rho-family G proteins with GTP which then bind and activate the associated Pak. In this application we characterize a novel guanine nucleotide exchange factor that is the corresponding binding partner of Pak4 and seems to show exchange factor activity specific for cdc42 (this protein has since been identified by Fukuhara et al. (1999)  Journal of Biological Chemistry  274(9):5868-5879).  
           [0007]    Besides Nef another essential HIV auxiliary protein, namely the unique HIV-encoded transcription factor Tat, has also been found to associate with cytoplasmic serine/threonine kinase activity (reviewed in Karn (1999)  Journal of Molecular Biology  293:235-254 and Taube et al. (1999) Virology 264:245-253). Tat recruits two cyclin-dependent-kinase-containing complexes (Cdk9/P-TEFb and Cdk7/TFIIH) to the HIV-LTR where by phosphorylating RNA polymerase 11 they contribute to transcriptional elongation. Until now these are the only Tat-associated cellular kinase activities to have been characterized.  
           [0008]    HIV-Tat also binds a cellular protein called Tip60 or 60 kD Tat-interacting protein (Kamine et al. (1996)  Virology  216:357-366). The sequence (SEQ ID NO:9 and SEQ ID NO:10) of this protein shows homology to known protein acetyl transferase enzymes, and indeed Tip60 has been shown to have acetyltransferase activity as assayed in vitro on histones (Kimura and Horikoshi (1998)  Genes to Cells  3:789-800). A good in vivo substrate for this enzyme has until now not been identified however.  
           [0009]    Accordingly, there remains a need for the elucidation of protein-protein binding interactions that regulate the activity of HIV-Tat and/or NEF. The identification of such interactions would be useful, for example, in screening for compounds that modulate the activity of Tat and/or NEF, and would enable the modulation of the activity of these proteins by modulating their protein-protein complex formation.  
         SUMMARY OF THE INVENTION  
         [0010]    In accordance with the present invention, it has now been demonstrated that HIV-Tat and HIV-Nef bind a complex of cellular signaling proteins including the serine/threonine kinase PAK4, the guanyl nucleotide exchange factor cdc42-GEF and the acetyl-transferase Tip60. Using an HIV-LTR transcriptional reporter assay we show that PAK4 and Cdc42/GEF synergize to activate Tat transcriptional activity, while Nef and Tip60 synergize to inactivate Tat. As effectors or modulators of the activity of the HIV auxiliary proteins, the cellular proteins PAK4, cdc42-GEF and Tip60 represent novel targets for the development of therapeutics directed against HIV. We also show that HIV-Nef is an in vivo substrate of Tip60 acetyl-transferase activity, providing a novel assay for the development of drugs that modulate Tip60 activity.  
           [0011]    The invention provides, in part, isolated DNA sequences encoding a novel PAK4 serine/threonine kinase, and a vector for expressing Cdc42-specific GEF (guanyl-nucleotide exchange factor), the vector comprising a DNA sequence encoding the same. Also provided are a method for producing PAK4 or Cdc42-GEF protein by the steps of transfecting a cell with a vector comprising a DNA sequence encoding PAK4 or Cdc42-GEF, and culturing the cell in order to express the desired vector.  
           [0012]    The invention also provides methods for modulating the transcriptional activity of HIV Tat protein by modulating the formation of a complex between Tat and at least one modulator complex comprising PAK4 and one or more binding proteins, and methods for identifying a compound that inhibits the transcriptional activity of HIV-Tat by determining whether the compound disrupts the formation of a complex including PAK4 and Cdc42-GEF, or whether the compound enhances the formation of a complex including PAK4, HIV-NEF, and Tip60. Also provided are methods of inhibiting the transcriptional activity of HIV-Tat using a compound that decreases or increases activity or expression of a HIV-Tat complex binding protein. Methods for modulating the activity of HIV-NEF, identifying a compound that modulates HIV-NEF acetylation by Tip60, and identifying a compound that modulates Tip60 acetyl-transferase activity are also provided. 
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0013]    [0013]FIG. 1 illustrates human multiple tissue Northern blot (Clontech) probed with PAK4 cDNA, revealing a ubiquitous 2.9 kb mRNA (liver signal visible on long exposure).  
         [0014]    [0014]FIG. 2A is a schematic of the structure of the PAK4 protein.  
         [0015]    [0015]FIG. 2B is a schematic of the structure of the GEF protein.  
         [0016]    [0016]FIG. 3 illustrates the epitope-tagged eukaryotic expression constructs used in co-immunoprecipitation experiments.  
         [0017]    [0017]FIG. 4A illustrates the co-precipitation of GST-PAK4 detected by anti-GST Western blot, showing that PAK4 binds GEF, Tat and Tip60 and Nef in vivo.  
         [0018]    [0018]FIG. 4B illustrates the co-precipitation of FLAG-GEF detected by anti-FLAG Western blot, showing that GEF specifically binds PAK4 but not PAK2 in vivo. GEF interacts with both the kinase domain and the amino-terminal regulatory domain of PAK4.  
         [0019]    [0019]FIG. 5A illustrates co-precipitation of FLAG-Tat detected by anti-FLAG Western blot, showing that HIV-Tat specifically binds PAK4 not PAK2 in vivo. Tat interacts with the amino-terminal regulatory domain, not the kinase domain, of PAK4.  
         [0020]    [0020]FIG. 5B illustrates the co-precipitation of FLAG-Nef detected by anti-FLAG Western blot. While an interaction between HIV-Nef and full-length PAK4 was not seen, a specific interaction between Nef and the amino-terminal regulatory domain of PAK4 (but not PAK2) was observed. The Nef-PAK4 interaction may occur in vivo at a point in the regulation of PAK4 activity when this binding domain is exposed.  
         [0021]    [0021]FIG. 6 illustrates co-precipitation of FLAG-tagged GEF, Nef and Tat detected by anti-FLAG Western blot, indicating the presence of distinct binding sites on PAK4 for Tat and Nef. Nef binds at the extreme amino terminus of PAK4, between amino acids 1 and 91 where the Cdc42-binding motif is also found, while Tat binds PAK4 between the Cdc42-binding motif and the kinase domain (between amino acids 93 and 290).  
         [0022]    [0022]FIG. 7A illustrates the co-precipitation of Cdc42 detected by anti-Cdc42 Western blot, showing that Cdc42 binds the amino terminus of PAK4 and that this binding is not disrupted by co-expression of Nef.  
         [0023]    [0023]FIG. 7B illustrates the co-precipitation of Rac detected by anti-Rac Western blot, showing that Rac does not strongly bind PAK4.  
         [0024]    [0024]FIG. 8A shows that over-expression of a truncated GEF protein (containing only the catalytic and pleckstrin homology domains) strongly activates Tat transcriptional activity (as measured using an HIV-LTR transcriptional reporter assay), but in a constitutive, unregulated fashion.  
         [0025]    [0025]FIG. 8B shows that over-expression of either a longer GEF protein (with a full carboxy-terminal tail) or the PAK4 protein does not stimulate Tat greatly, but when both are co-expressed with Tat they synergize to strongly activate Tat transcriptional activity.  
         [0026]    [0026]FIG. 9 shows that activation of Tat by PAK4 is strongly inhibited by Rac, and strongly enhanced by Cdc42. Expression of dominant-negative Rac (RacNl7) or even wild-type Rac blocks PAK4 activation of Tat and even basal Tat activity. Wild-type Cdc42 or an activated mutation of Cdc42 (61L) strongly enhance activation of Tat by PAK4, consistent with Cdc42 (but not Rac) being the substrate of GEF guanyl nucleotide exchange factor activity and mediating its effects on Tat.  
         [0027]    [0027]FIG. 10 shows Nef acetylation by Tip60 in vivo in COS cells transfected with FLAF-Nef and additionally transfected either with or without Tip60. Nef was immunoprecipitated with anti-FLAG followed by Western blot with anti-acetylated lysine antibody (Catalog #9441, Cell Signaling Technology, Inc.). Amount of Nef protein is the same in the two lanes (data not shown).  
         [0028]    [0028]FIG. 11 is a comparison of the sequence of human (SEQ ID NO:1) and Drosophila PAK4 (SEQ ID NO:2). 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0029]    The present invention is based, in part, on the identification and sequencing of a novel serine/threonine kinase, termed “PAK4” herein. The invention is also based, in part, on the discovery of HIV-Tat and HIV-NEF complex formation with PAK4, Cdc42-GEF (guanyl-nucleotide exchange factor), and/or the acetyl-transferase Tip60, which complex formations regulate the activity of HIV-Tat, HIV-NEF and/or Tip60, respectively. The identification of these interactions provides a novel means for identifying compounds that modulate the activity of these enzymes, and or for using such compounds to modulate HIV transcriptional enzymatic activity, as further described below. All references cited are hereby incorporated herein by reference.  
         [0030]    PAK4 Characterization &amp; Isolated PAK4 DNA  
         [0031]    PAK4 was initially identified as an expressed sequence tag in the GenBank database (Genbank accession number T83145, IMAGE clone 110764) during a search for novel members of this interesting kinase family. This expressed sequence tag was then used to screen a human SK-N-MC cell cDNA library in order to isolate a full length cDNA (GenBank accession number AF005046; SEQ ID NO:3 and SEQ ID NO:4), as further described in Example 1.  
         [0032]    A Drosophila homolog of PAK4 was characterized at the same time by searching the Berkeley Drosophila Genome Database (www.fruitfly.org) with the sequence of the human Pak4 cDNA. A Drosophila cDNA clone (LD05866) that turned out to be full length was obtained from the Berkeley Drosophila Genome Project (GenBank Accession No. AF031517; SEQ ID NO:5 and SEQ ID NO:6; since published by Melzig et al. (1998)  Current Biology  8:1223-1226), as further described in Example 1. FIG. 11 compares the sequence of the human and Drosophila Pak4 proteins, highlighting domains of functional significance including the kinase domain and G-protein binding domain.  
         [0033]    The invention provides, therefore, in part, isolated DNA molecules encoding the novel PAK4 kinase. In a preferred embodiment, the invention provides an isolated DNA sequence encoding PAK4 serine/threonine kinase, wherein the sequence comprises SEQ ID NO: 1 or conservative mutants or variants thereof. As used herein, “conservative mutants or variants” of a nucleotide or peptide sequence means sequences varying from the specific nucleotide or peptide sequence due to the degeneracy of the genetic code or to point mutations or variances in sequence that due not affect the activity of the encoded protein. In another embodiment, the invention provides a method for producing PAK4 protein comprising the steps of: (a) transfecting a cell with a vector comprising a DNA sequence encoding PAK4, and (b) culturing said cell under conditions suitable for the expression of the desired vector. In another embodiment, the invention provides recombinant PAK4 protein produced by this method, the protein comprising the amino acid sequence of SEQ ID NO: 4 or conservative mutants or variants thereof.  
         [0034]    Cdc42-GEF Characterization and Isolated Cdc42-GEF DNA  
         [0035]    In order to help characterize the cellular function of the novel PAK4 kinase disclosed herein, a yeast two-hybrid screen of a human brain library was carried out in order to identify potential interacting proteins, as further described in Example II. During this screen, using the human PAK4 open reading frame as bait, a carboxy-terminal fragment of a putative rho-family guanyl nucleotide exchange factor (GEF) previously known only as a cDNA sequence (GenBank Accession No. AB002378; SEQ ID NO:7 and SEQ ID NO:8)) was identified. As described below, Cdc42-GEF complexes with HIV-Tat and modulates its activity.  
         [0036]    Accordingly, the invention also provides, in part, isolated DNA molecules encoding Cdc42-GEF protein, and expression vectors containing the same. In a preferred embodiment, there is provided a vector for expressing Cdc42-specific GEF (guanyl-nucleotide exchange factor), said vector comprising a DNA sequence selected from the group consisting of SEQ ID NO: 7, residues 640 to 1105 of SEQ ID NO: 7, residues 640 to 1522 of SEQ ID NO: 7, and conservative mutants or variants thereof. As further described in Example II below, the short form (residues 640-1522) contains the putative catalytic domain and pleckstrin homology (PH) domains (FIGS. 2A and 2B), and the long form (residues 640-1522) contains these domains plus the 400 amino acid residues C-terminal of the PH domain. The GEF long form modestly activates Tat, but dramatically activated Tat in combination with PAK4, while GEF short form constitutively and dramatically activates Tat (FIG. 3), as further described in Example 3.  
         [0037]    In another embodiment, the invention provides a method for producing Cdc42-GEF protein, the method comprising the steps of: (a) transfecting a cell with a vector comprising a Cdc42-GEF DNA sequence, and (b) culturing said cell under conditions suitable for the expression of the desired vector. The invention also provides a recombinant Cdc42-GEF protein produced by this method, wherein said protein comprises the amino acid sequence of SEQ ID NO: 8 or conservative mutants or variants thereof.  
         [0038]    Co-immunoprecipitation Experiments  
         [0039]    In order to confirm the PAK4 protein interaction complexes identified in yeast (see Example 1), epitope-tagged eukaryotic expression constructs were made so that protein-protein interactions and complexes could be further tested in vivo by means of co-immunoprecipitation experiments, as further described in Example 2. As part of this screening, given the interaction with Tip60, the interaction of PAK4 with HIV-Tat itself was examined. Further, given the potential HIV connection and mindful of the literature reporting HIV-Nef interacting with a PAK-family kinase (Trono and Wang (1997)  Chemistry and Biology  4:13-15), the possible interactions of HIV-Nef with PAK4 was examined. See Example 2.  
         [0040]    The interactions between PAK4 and the cellular proteins Tip60 and GEF were confirmed by co-immunoprecipitation of the proteins expressed in COS cells (FIG. 4A); see Example 2. The binding of GEF was specific for PAK4 (i.e. GEF was not co-precipitated by PAK2 (FIG. 4B). The interaction of GEF with PAK4 parallels the recent observation of binding of the PIX family of guanyl nucleotide exchange factors to other members of the PAK family (Manser et al. (1998)  Mol. Cell  1:183-192).  
         [0041]    When the possibility of interactions between PAK4 and the HIV accessory proteins Tat and Nef was examined, full-length PAK4 was found to clearly pull down Tat (FIGS. 5A and 5B). In order to localize the Tat-binding region within the PAK4 protein, amino- and carboxy-terminal halves of PAK4 were expressed separately. Co-immunoprecipitation experiments with these PAK4 deletion constructs showed strong binding of both Tat and Nef to the amino-terminal, putative regulatory half of PAK4 (amino acids residues 1-290) but not the carboxy-terminal kinase domain; furthermore this interaction was specific for PAK4, as neither Nef nor Tat bound corresponding PAK2 expression constructs (FIGS. 5A and 5B). See Example 2.  
         [0042]    When the specificity of PAK4 for Cdc42 vs. Rac was examined by co-immunoprecipitation of PAK4 co-expressed in COS cells with either of the two small G proteins, it was observed that PAK4 binds Cdc42 not Rac, and furthermore this interaction was not disrupted by co-expressing Nef (FIGS. 7A and 7B).  
         [0043]    The regulation of Tat transcriptional activity by PAK4 and PAK4-associated proteins was further examined, as described in Example 3, in COS cells using a reporter construct containing luciferase gene under the control of the HIV-LTR promoter. These experiments indicate that PAK4 and Cdc42-GEF form a protein complex with HIV Tat and modulate its activity.  
         [0044]    The regulation of HIV-Nef activity via acetylation by Tip60 and associated binding proteins was further examined, as described in Example 3, in COS cells expressing PAK4, GEF, Tat, and Nef, with or without Tip60. These experiments indicate that HIV-Nef is acetylated in the presence of Tip60. None of the other proteins tested showed acetylation by Tip60, as assayed by Western blot of cell lysates using an antibody that recognizes acetylated lysine regardless of amino acid context.  
         [0045]    Modulation of Tat Activity &amp; Screening Methods  
         [0046]    The identification of protein-protein interactions and complexes disclosed herein provides a novel means for modulating the activity of HIV transcriptional proteins and screening for compounds that modulate (i.e. activate or inhibit) the activity of these proteins.  
         [0047]    Thus, in one embodiment, the invention provides a method for modulating the transcriptional activity of human immunodeficiency virus (HIV) Tat protein, said method comprising modulating the formation of a complex between Tat and at least one modulator complex comprising (i) the serine/threonine kinase PAK4 and the guanyl nucleotide exchange-factor Cdc42-GEF or (ii) PAK4, HIV-NEF, and the acetyl-transferase Tip60. In a preferred embodiment, the modulator complex comprises PAK4/Cdc42-GEF, and the inhibition of formation of a complex between Tat and said modulator complex decreases the transcriptional activity of Tat. In another preferred embodiment, the formation of the complex between Tat and said modulator complex is inhibited by contacting a Tat-expressing cell or cellular preparation with at least one compound that decreases the activity or expression of PAK4 and/or Cdc42-GEF. In another preferred embodiment, the modulator complex comprises PAK4/HIV-NEF/Tip60, and the formation of a complex between Tat and said modulator complex decreases the transcriptional activity of Tat. In still another preferred embodiment, the formation of the complex between Tat and said modulator complex is induced by contacting a Tat-expressing cell or cellular preparation with at least one compound that alters the activity or expression of PAK4 and/or HIV-NEF, and/or Tip60. In a preferred embodiments, the compound is an inhibitor of PAK4 kinase activity or an inhibitor of the GTP exchange factor activity of cdc42-GEF.  
         [0048]    The inhibition of a complex between Tat and the PAK4/cdc42-GEF modulator complex decreases the transcriptional activity of Tat, and the inhibition of a complex between Tat and the PAK4/Nef/Tip60 modulator complex increases the transcriptional activity of Tat. As described herein, these characteristics can be used in the search for novel drug candidates against HIV, with the inhibition of the transcriptional activity of Tat as a target. In many drug screening programs which test libraries of compounds and natural extracts, high throughput assays are desirable in order to maximize the number of compounds surveyed in a given period of time. Assays which are performed in cell-free systems, such as may be derived with purified or semi-purified proteins, are often preferred as “primary” screens in that they can be generated to permit rapid development and relatively easy detection of an alteration in a molecular target which is mediated by a test compound. Moreover, the effects of cellular toxicity and/or bioavailability of the test compound can be generally ignored in the in vitro system, the assay instead being focused primarily on the effect of the drug on the molecular target. The cell-free assay involving immunoprecipitation as described below is suitable for a high throughput format that can be designed for robotic automation.  
         [0049]    In another embodiment, therefore, the invention provides a method for identifying a compound (e.g. a drug candidate) that inhibits the transcriptional activity of HIV-Tat, said method comprising the steps of: reacting said compound with a complex comprising (i) PAK4/Cdc42-GEF or (ii) HIV-Tat/PAK4/Cdc42-GEF; and determining whether said complex of step (a) is disrupted, wherein said compound is identified as an inhibitor of HIV-Tat transcriptional activity if said complex is disrupted. In preferred embodiments, the complex of step (a) is present in a cellular extract, and the determination of step (b) is accomplished by immunoprecipitation. In another preferred embodiment, the method further comprises the step of (c) confirming that the compound inhibits the in vivo transcriptional activity of Tat by reacting said compound with a cell or cellular preparation comprising a Tat transcriptional reporter. In a preferred embodiment, the transcriptional reporter comprises luciferase activity.  
         [0050]    A drug candidate, for example, may be assessed for its capability of inhibiting the transcriptional activity of human immunodeficiency virus (HIV) Tat protein by testing on a cellular extract containing a complex of (i) PAK4/cdc42-GEF or (ii) HIV-Tat/PAK4/cdc42-GEF. The complex is immunoprecipitated with an antibody immobilized, for example, on beads. The immobilized antibody reacts with one of the proteins in the complex, or if one of the proteins is tagged, the antibody also can react against the protein tag. The complex bound to the antibody is released from the antibody after precipitation and washing of the beads by, for example, boiling in SDS-containing buffer. The amount of complex formed in the cellular extract then is determined by analyzing the amount of protein released from the antibody, for example by separating the proteins by SDS-PAGE and probing by Western blot. The influence of a drug candidate on the formation of said complexes is measured against a control incubation that does not contain the drug candidate investigated. The capability of a drug candidate to inhibit the transcriptional activity of human immunodeficiency virus (HIV) Tat protein is measured by its ability to disrupt said complexes.  
         [0051]    In another embodiment, the invention provides a method for identifying a compound that inhibits the transcriptional activity of HIV-Tat, the method comprising the steps of: (a) reacting said compound with a mixture comprising (i) PAK4, HIV-NEF, and Tip60 or (ii) HIV-Tat, PAK4, HIV-NEF, and Tip60; and (b) determining whether said compound enhances the formation of a complex comprising (i) PAK4/HIV-NEF/Tip60 or (ii) HIV-Tat/PAK4/HIV-NEF/Tip60, wherein said compound is identified as an inhibitor of HIV-Tat transcriptional activity if the formation of a complex in step(b) is enhanced. In preferred embodiments, the mixture of-step (a) is present in a cellular extract, and the determination of step (b) is accomplished by immunoprecipitation. In another preferred embodiment, the determination of step (b) is accomplished by comparing complex formation to the level of complex formation in a control sample. In still another preferred embodiment, the method further comprises the step of (c) confirming that the compound inhibits the in vivo transcriptional activity of Tat by reacting said compound with a cell or cellular preparation comprising a Tat transcriptional reporter. In a preferred embodiment, the transcriptional reporter comprises luciferase activity.  
         [0052]    For example, a drug candidate may be assessed for its capability of inhibiting the transcriptional activity of human immunodeficiency virus (HIV) Tat protein by testing on a cellular extract containing a complex of (i) PAK4/HIV-Nef/Tip60 or (ii) HIV-Tat/PAK4/HIV-Nef/Tip60. The complex is immunoprecipitated with an antibody immobilized, for example, on beads. The immobilized antibody reacts with one of the proteins in the complex, or if one of the proteins is tagged, the antibody also can react against the protein tag. The complex bound to the antibody is released from the antibody after precipitation and washing of the beads by, for example, boiling in SDS-containing buffer. The amount of complex formed in the cellular extract then is determined by analyzing the amount of protein released from the antibody, for example by separating the proteins by SDS-PAGE and probing by Western blot. The influence of a drug candidate on the formation of said complexes is measured against a control incubation that does not contain the drug candidate investigated. The capability of a drug candidate to inhibit the transcriptional activity of human immunodeficiency virus (HIV) Tat protein is measured by its ability to enhance the formation of said complexes.  
         [0053]    Other techniques for immobilizing proteins on matrices are also available for use in the subject immunoprecipitation assays. For example, glutathione-S-transferase/PAK4 (GST/PAK4) fusion proteins can be adsorbed onto glutathione sepharose beads (Amersham Pharmacia or Sigma Chemical) or glutathione derivatized microtitre plates, which are then combined with the cdc42-GEF, Nef or Tat polypeptides. In another example, either of the PAK4, cdc42-GEF, Nef or Tat proteins can be immobilized utilizing conjugation of biotin and streptavidin. For instance, biotinylated PAK4 molecules can be prepared from biotin-NHS (N-hydroxy-succinimide) using techniques well known in the art (e.g., biotinylation kit, Pierce Chemicals, Rockford, Ill.), and immobilized in the wells of streptavidin-coated 96 well plates (Pierce Chemical). Alternatively, antibodies reactive with PAK4 but which do not interfere with binding of cdc42-GEF, Nef and Tat can be derivatized to the wells of the plate, and the PAK4 trapped in the wells by antibody conjugation. As above, preparations of cdc42-GEF, Nef or Tat polypeptides and a test compound are incubated in the PAK4-presenting wells of the plate, and the amount of complex between PAK4 and and one or more of the above proteins trapped in the well can be quantitated. Exemplary methods for detecting such complexes, in addition to those described above for the GST-immobilized complexes, include immunodetection of complexes using antibodies reactive with the cdc42-GEF, Nef or Tat polypeptides, or which are reactive with the PAK4 protein and compete for binding with the cdc42-GEF, Nef or Tat polypeptides; as well as enzyme-linked assays (see below).  
         [0054]    Complex formation between the PAK4 polypeptide and a cdc42-GEF polypeptide, a Tip60 polypeptide, a Nef polypeptide or a Tat polypeptide may be detected by a variety of techniques. In the above outline of the immunoprecipitation technique, an example was given where the proteins are separated by SDS-PAGE and probed by Western blot. Alternatively, modulation of the formation of complexes can be quantitated using, for example, detectably labelled proteins such as radiolabelled (e.g.  32 P,  35 S,  14 C or  3 H) cdc42-GEF, Nef, or Tat polypeptides. Following incubation, the beads or microtiter plates containing immobilized PAK4 polypeptides are washed to remove any unbound cdc42-GEF, Nef or Tat polypeptides, and the matrix immobilized radiolabel determined directly (e.g. beads placed in scintilant), or in the supernatant after the complexes of PAK4 with cdc42-GEF, Nef or Tat proteins are subsequently dissociated.  
         [0055]    In yet another alternative, enzyme-linked assays are used which rely on detecting an enzymatic activity associated with the cdc42-GEF, Nef or Tat polypeptides (instead of the intrinsic activity of the cdc42-GEF, Nef or Tat polypeptides). In the instance of the latter, the enzyme can be chemically conjugated or provided as a fusion protein with cdc42-GEF, Nef or Tat polypeptides. To illustrate, the cdc42-GEF, Nef or Tat polypeptides can be chemically cross-linked or genetically fused with horseradish peroxidase, and the amount of cdc42-GEF, Nef or Tat polypeptides trapped in the complex can be assessed with a chromogenic substrate of the enzyme, e.g. 3,3′-diamino-benzadine terahydrochloride or 4-chloro-1-napthol. Likewise, a fusion protein comprising the cdc42-GEF, Nef or Tat polypeptides and glutathione-S-transferase can be provided, and complex formation quantitated by detecting the GST activity using 1-chloro-2,4-dinitrobenzene (Habig et al (1974)  J Biol Chem  249:7130). In yet another application, proteins may be fluorescently labelled (e.g. FITC), and their presence in the complex may then be measured in a fluorometer. Alternatively, the presence of a complex may be assayed by means of fluorescence resonance energy transfer (FRET), whereby one member of the complex is labeled with a fluorescence donor molecule and another labeled with a fluorescence acceptor molecule, such that in the intact complex fluorescence signal is emitted, but on disruption of the complex energy transfer is interrupted and the signal is lost. FRET may be carried out according to protocols well known to those in the art.  
         [0056]    In still another embodiment, the invention provides a method for inhibiting the transcriptional activity of HIV-Tat, the method comprising contacting a HIV-Tat-expressing cell with at least one compound selected from the group consisting of:  
         [0057]    (i) a compound that decreases activity or expression of PAK4;  
         [0058]    (ii) a compound that decreases activity or expression of Cdc42-GEF;  
         [0059]    (iii) a compound that increases activity or expression of HIV-NEF; and  
         [0060]    (iv) a compound that increases activity or expression of Tip60.  
         [0061]    Since the formation of the complexes of (i) PAK4/cdc42-GEF or (ii) HIV-Tat/ PAK4/cdc42-GEF or (iii) PAK4/HIV-Nef/Tip6O or (iv) HIV-Tat/PAK4/HIV-Nef/Tip60, modulate the transcriptional activity of Tat, Tat activity may be regulated by a drug candidate that influences the expression or activity of the proteins involved in the complex. The action on a cell by a drug candidate that decreases the expression of (i) PAK4 and/or (ii) cdc42-GEF or that increases the expression of (i) HIV-Nef and/or (ii) Tip60 will lead to inhibition of the transcriptional activity of Tat as well.  
         [0062]    For example, a drug candidate may be added to cells for a certain amount of time before harvesting, a cell extract is made, and the cell extract is tested for the amount of said proteins expressed. The determination of the amount of said proteins is performed, for example, by separating the proteins by SDS-PAGE and probing by Western blot. The influence of a drug candidate on the expression of said proteins is measured against a control incubation that does not contain the drug candidate investigated. The capability of a drug candidate to inhibit the transcriptional activity of human immunodeficiency virus (HIV) Tat protein is measured by its ability to decrease the expression of (i) PAK4 and/or (ii) cdc42-GEF or to increase the expression of (i) HIV-Nef and/or (ii) Tip60. Alternatively, expression may be monitored in a cell-based, high-throughput assay system (e.g. High Content Screening (HCS)), such as HCS sytems offered by Cellomics, Inc.  
         [0063]    A drug candidate identified in any of the above assays is further confirmed to inhibit transcriptional activity of Tat in vivo. In this assay, the drug candidate is reacted with a cell or cellular preparation comprising a Tat transcriptional reporter. In a preferred embodiment of this assay, the Tat transcriptional reporter comprises luciferase activity which is measured, for example, using the Roche Biochemicals luciferase assay kit and a Victor2 luminometer (Perkin Elmer Life Sciences).  
         [0064]    In addition to the above assays, the present invention further contemplates screening the drug candidates against Tat selected with the above methods in assays that may serve as an intermediate step for clinical trials on humans. Suitable for that purpose, for example, may be a microtiter assay which measures the ability of selected compounds to inhibit HIV-induced cell killing as well as the toxicity of the test compounds to host cells. The basic assay involves infection of CEM-SS cells or other human cells with virus in the presence of the test compound. Quantitation may be performed, for example, spectrophotometrically using the tetrazolium dye MTS (Cell Titer; Promega) which is converted to a soluble, colored formazan product by mitochondrial enzymes present in metabolically active cells at six days post-infection. Confirmatory assays may include, for example, reverse transcriptase, p24 and infectious virus assays, as well as macroscopic and microscopic observation of test wells.  
         [0065]    Finally, clinical trials may be performed on drug candidates that have tested positive in all previous biochemical and cell-based assays. The response of human patients to the drug candidates may be monitored by analyzing cells extracted from the patients in the CTL killing assay, the limited dilution analysis (LDA), or by newer methods such as ELISpot and Intracellular cytokine staining (ICC).  
         [0066]    Modulation of HIV-NEF and Tip60 Activity &amp; Screening Methods  
         [0067]    The identification of HIV-NEF and Tip60 protein complex formation disclosed herein also provides novel methods for the modulation of the activity of either of these enzymes by modulation of such complex formation.  
         [0068]    Thus, in one embodiment, the invention further provides a method for modulating the activity of HIV-NEF, the method comprising contacting a HIV-NEF-expressing cell with at least one compound that modulates the acetyl-transferase activity of Tip60. In a preferred embodiment, the compound increases the activity or expression of Tip60. In another preferred embodiment, the compound decreases the activity or expression of Tip60.  
         [0069]    In another embodiment, the invention provides a method for identifying a compound that modulates HIV-NEF acetylation by the acetyl-transferase Tip60, the method comprising the steps of (a) reacting said compound with a mixture comprising HIV-NEF and Tip60; and (b) determining whether said compound inhibits or enhances the level of acetylation of HIV-NEF, wherein said compound is identified as a modulator of HIV-NEF acetylation by Tip60 if the level of acetylation in step(b) is inhibited or enhanced.  
         [0070]    In still another embodiment, the invention provides a method for identifying a compound that modulates Tip60 acetyl-transferase activity, the method comprising the steps of: (a) reacting said compound with a mixture comprising HIV-NEF and Tip60; and (b) determining whether said compound inhibits or enhances the level of acetylation of HIV-NEF, wherein said compound is identified as a modulator of Tip60 acetyl-transferase activity if the level of acetylation in step(b) is inhibited or enhanced. In a preferred embodiment, the determination of step (b) is accomplished by comparing the level of acetylation of HIV-NEF to the level of acetylation in a control sample. The compound may thus either decrease or increase the activity and/or expression of Tip60. Tip60 acetylates HIV-Nef, and that acetylation in turn may either increase or decrease the activity of HIV-Nef.  
         [0071]    As disclosed herein, HIV-Nef has now been shown to be a substrate of the acetyl-transferase activity of Tip60. Thus, for example, the method of the invention may be employed to identify a drug candidate that modulates Tip60 acetyl-transferase activity. In many drug screening programs which test libraries of compounds and natural extracts, high throughput assays are desirable in order to maximize the number of compounds surveyed in a given period of time. Assays which are performed in cell-free systems, such as may be derived with purified or semi-purified proteins, are often preferred as “primary” screens in that they can be generated to permit rapid development and relatively easy detection of an alteration in a molecular target which is mediated by a test compound. Moreover, the effects of cellular toxicity and/or bioavailability of the test compound can be generally ignored in the in vitro system, the assay instead being focused primarily on the effect of the drug on the molecular target. The cell-free assay involving reaction of Tip60 with HIV-Nef as described below is suitable for a high throughput format that can be designed for robotic automation.  
         [0072]    In a preferred embodiment, a drug candidate to be screened for its ability to modulate the acetyl-transferase activity of Tip60 is reacted with a mixture comprising HIV-Nef and Tip60. The proteins may be present in a semi-purified or in a purified form. The amount of acetylation of HIV-Nef then is established, for example, by separating the proteins by SDS-PAGE and probing by Western blot with an antibody against acetylated lysine, such as Cell Signaling Technology, Inc. catalog # 9941. The influence of a drug candidate on the acetylation of HIV-Nef, which may be either inhibitory or enhancing, then is established by comparing the level of acetylation of HIV-Nef to the level of acetylation in a control sample.  
         [0073]    The following Examples are provided only to further illustrate the invention, and are not intended to limit its scope, except as provided in the claims appended hereto. The present invention encompasses modifications and variations of the methods taught herein which would be obvious to one of ordinary skill in the art.  
       EXAMPLE 1  
       [0074]    Identification of PAK4 and its Binding Partners  
         [0075]    PAK4 kinase was initially found as an expressed sequence tag in the GenBank database (Genbank accession number T83145, IMAGE clone 110764) during a search for novel members of this interesting kinase family. This expressed sequence tag was used to screen a human SK-N-MC cell cDNA library in the vector pCDNA1 (Invitrogen) and isolate a full length cDNA (GenBank accession number AF005046; SEQ ID NO:3 and SEQ ID NO:4). Northern analysis using this cDNA to probe a human multiple tissue northern blot (Clontech) indicates that expression of this gene is ubiquitous (FIG. 1). A BAC human genomic DNA clone (14B11) containing this gene was identified by Research Genetics, Inc. (Huntsville, Ala.), and mapped by them to human chromosome 19q13.  
         [0076]    A Drosophila homolog of PAK4 was also characterized at the same time by searching the Berkeley Drosophila Genome Database (www.fruitfly.org) with the sequence of the human Pak4 cDNA. A Drosophila cDNA clone (LD05866) that turned out to be full length was obtained from the Berkeley Drosophila Genome Project (GenBank Accession No. AF031517; SEQ ID NO:5 and SEQ ID NO:6; since published by Melzig et al. (1998)  Current Biology  8:1223-1226). FIG. 11 compares the sequence of the human and Drosophila Pak4 proteins, highlighting domains of functional significance including the kinase domain and G-protein binding domain. A Drosophila genomic DNA clone containing this gene was identified using the Drosophila cDNA to screen an array of BAC clones provided by the Berkeley Drosophila Genome Project; these previously mapped clones gave a chromosomal map position for the Drosophila gene of 14E on the X chromosome.  
         [0077]    Initial Identification of Interacting Proteins  
         [0078]    In order to help characterize the cellular function of this novel kinase, a yeast two-hybrid screen of a human brain library in vector pACT2 was performed to identify potential interacting proteins (Matchmaker GAL4 Two-hybrid system, Clontech). Using the human PAK4 open reading frame cloned into the vector pAS2-1 as bait, a carboxy-terminal fragment of a putative rho-family guanyl nucleotide exchange factor (GEF) previously known only as a cDNA sequence (GenBank Accession No. AB002378; SEQ ID NO:7 and SEQ ID NO:8)) was identified as a complexing protein. Using the amino-terminal half of PAK4 (amino acids 1 to 313, lacking the kinase domain) cloned into the vector pAS2-1 as bait, a putative acetyl-transferase, called Tip60 because it was initially characterized as an HIV-Tat-interacting protein (Kamine et al. (1996)  Virology  216:357-366), was identified as a complexing protein. This enzyme is also evolutionarily conserved in Drosophila where it is called Mof and implicated in dosage compensation (Hilfiker et al. (1997)  EMBO J.  16(8):2054-2060).  
       EXAMPLE 2  
       [0079]    In vivo Protein-Protein Binding Interactions of HIV-Tat and -Nef  
         [0080]    In order to confirm the PAK4 protein interactions identified in yeast, epitope-tagged eukaryotic expression constructs were made so that protein-protein interactions could be further tested in vivo by means of co-immunoprecipitation experiments. PAK4 and various PAK4 deletions were expressed as GST fusion proteins using a eukaryotic expression vector kindly provided by John Kyriakis, where an amino-terminal GST fusion of the desired protein is expressed under the control of the strong constitutive EF-1α promoter (Mizushima and Nagata (1990)  Nucleic Acids Research  18(17):5322). Tip60 and GEF were expressed as FLAG-tagged proteins after cloning into the pFLAG-CMV-2 vector from Sigma-Aldrich (FIGS. 2A, 2B, and  3 ).  
         [0081]    Further, given the interaction with Tip60, the possibility that PAK4 might interact with HIV-Tat itself was tested. The HIV-Tat open reading frame was generated by gene synthesis (Stemmer et al. (1995)  Gene  164(1):49-53) using the Tat amino acid sequence from the SF2 isolate of HIV-1 (Genbank accession number AAB59879) converted to DNA sequence using optimal human codon usage tables and the backtranslate program from the Wisconsin Package of the Genetics Computer Group (GCG; Madison, Wis.). The Tat open reading frame was then cloned into the vector pFLAG-CMV-2 in order to express FLAG-tagged protein. Given the potential HIV connection and mindful of the literature reporting HIV-Nef interacting with a PAK-family kinase (Trono and Wang (1997)  Chemistry and Biology  4:13-15), a FLAG-tagged Nef (Genbank accession number AAC68849) was also synthesized and expressed in order to investigate a possible interaction with PAK4.  
         [0082]    COS cells seeded into 6-well plates were transfected using Fugene-6 (Roche Biochemicals), and various GST- and Flag-tagged proteins expressed or coexpressed for 4248 hrs. Cells were washed with PBS, then 1 ml of the following buffer was added:  
         [0083]    20 mM Tris/HCI pH 7.5  
         [0084]    50 mM NaCl  
         [0085]    1 mM EDTA  
         [0086]    1 mM EGTA  
         [0087]    0.1% Triton X-100  
         [0088]    plus the following phosphatase inhibitors:  
         [0089]    1 mM Sodium vanadate  
         [0090]    2.5 mM sodium pyrophosphate  
         [0091]    1 mM β-Glycerol-phosphate  
         [0092]    plus protease inhibitors (Roche Biochemicals Complete cocktail).  
         [0093]    Cells were lysed by passing through a 27 gauge needle four times, then spun for 10 minutes at maximum speed in an Eppendorf microcentrifuge. Anti-Flag co-immunoprecipitation was performed by adding 1 μg of antibody to 400 μl of the supernatant from the cell lysate, followed by overnight incubation at 4° C. The next day 20 μl protein A sepharose (or 20 μl of glutathione sepharose in the case of GST-pulldowns) were added, incubated for one hour on a rotator at 4° C., then washed three times with 500 μl buffer. Beads were boiled in SDS-sample buffer, and proteins in the sample analyzed by SDS-PAGE and western blotting.  
         [0094]    In the case of pulldowns of GST-fusions of PAK4, PAK4 ΔN, PAK4 ΔC, and PAK2 using glutathione sepharose, the co-precipitation of Flag-tagged protein partners was detected by Western blot with anti-Flag antibody (Kodak). In the case of immunoprecipitation of Flag-tagged gef, tip 60, tat, and nef with M5 anti-Flag antibody and protein A-sepharose, co-precipitated GST-fusion PAK proteins were detected by Western blot with anti-GST antibody.  
         [0095]    The interactions (i.e. complexes) between PAK4 and the cellular proteins Tip60 and GEF were confirmed by co-immunoprecipitation of the proteins expressed in COS cells (FIG. 4A). The binding of GEF was specific for PAK4 (i.e. GEF was not co-precipitated by PAK2 (FIG. 4B)), and disrupted by co-expression of Cdc42 or Rac. The interaction of GEF with PAK4 parallels the recent observation of binding of the PIX family of guanyl nucleotide exchange factors to other members of the PAK family (Manser et al. (1998)  Mol. Cell  1:183-192).  
         [0096]    When the possibility of interactions between PAK4 and the HIV accessory proteins Tat and Nef was examined, full-length PAK4 was found to clearly pull down Tat but not Nef (FIGS. 5A and 5B). In order to localize the Tat-binding region within the PAK4 protein, amino- and carboxy-terminal halves of PAK4 were expressed separately. Co-immunoprecipitation experiments with these PAK4 deletion constructs showed strong binding of both Tat and Nef to the amino-terminal, putative regulatory half of PAK4 (amino acids residues 1-290) but not the carboxy-terminal kinase domain; furthermore this interaction was specific for PAK4, as neither Nef nor Tat bound corresponding PAK2 expression constructs (FIGS. 5A and 5B).  
         [0097]    The amino-terminal, putative regulatory half of PAK4 (amino acids residues 1-290) was then further subdivided. PAK4-Na spans residues 1-91, containing a consensus nuclear localization signal and a consensus Rac/Cdc42-binding domain (FIGS. 2A and 2B). PAK4-Nb spans residues 93-290, in between the Rac/Cdc42-binding domain and the kinase domain, a region of no recognizable sequence motifs. This division clearly separated the Nef and Tat binding domains on PAK4, with Nef binding PAK4-Na with its Rac/Cdc42-binding domain, and Tat binding PAK4-Nb (FIG. 6). When the specificity of PAK4 for Cdc42 vs. Rac was examined by co-immunoprecipitation of PAK4 co-expressed in COS cells with either of the two small G proteins, it was observed that PAK4 binds Cdc42 not Rac, and furthermore this interaction was not disrupted by co-expressing Nef (FIGS. 7A and 7B).  
       EXAMPLE 3  
       [0098]    Regulation of Tat Transcriptional Activity by PAK4 and PAK4-associated Proteins  
         [0099]    A Tat transcriptional reporter construct containing luciferase under the control of the HIV-LTR promoter was kindly provided by Ben Berkhout (Verhoef et al. (1997)  Nucleic Acids Res.  25(3):496-502). Tat and the HIV-LTR-Luc reporter were co-transfected in COS cells along with PAK4 and PAK4-associated proteins, and luciferase activity assayed in cell lysates 30 hours post-transfection using the Roche Biochemicals luciferase assay kit and a Victor2 luminometer (Perkin Elmer Life Sciences).  
         [0100]    Co-expressing PAK4 with Tat modestly stimulated Tat activity as assayed by the HIV-LTR-Luc transcriptional reporter assay (FIGS. 8A and 8B). Two GEF expression constructs were also tested: a short form (residues 640-1105 of SEQ ID NO: 7) containing the putative catalytic domain and pleckstrin homology (PH) domain (FIGS. 2A and 2B), and a long form (residues 640-1522 of SEQ ID NO: 7) that had these domains plus the 400 amino acid residues C-terminal of the PH domain. The GEF long form modestly activated Tat, but dramatically activated Tat in combination with PAK4. GEF short form constitutively and dramatically activated Tat (FIG. 3). Cdc42 strongly activated Tat, and an activated mutation of Cdc42 (61L) stimulated Tat even more strongly, while Rac consistently inhibited Tat activity (FIG. 9).  
         [0101]    Acetylation of HIV-Nef by Tip60  
         [0102]    An anti-acetylated lysine antibody from Cell Signaling Technology, Inc. (catalog #9441) was used to look for a potential in vivo substrate of Tip60. PAK4, GEF, Tat and Nef were all expressed in COS cells with or without Tip60, then immunoprecipitated via their respective epitope tags and tested by anti-acetylated lysine western blot for acetylation induced by Tip60 co-expression. HIV-Nef demonstrated a high level of lysine acetylation in the presence of Tip60 (FIG. 10). Tip60 also showed auto-acetylation. None of the other proteins tested showed acetylation (data not shown), at least as detected by this antibody.  
     
       
       
         1 
         
           
             10  
           
           
             1  
             588  
             PRT  
             Homo sapiens  
           
            1 

Met Phe Gly Lys Arg Lys Lys Arg Val Glu Ile Ser Ala Pro Ser Asn 
  1               5                  10                  15 

Phe Glu His Arg Val His Thr Gly Phe Asp Gln His Glu Gln Lys Phe 
             20                  25                  30 

Thr Gly Leu Pro Arg Gln Trp Gln Ser Leu Ile Glu Glu Ser Ala Arg 
         35                  40                  45 

Arg Pro Lys Pro Leu Val Asp Pro Ala Cys Ile Thr Ser Ile Gln Pro 
     50                  55                  60 

Gly Ala Pro Lys Thr Ile Val Arg Gly Ser Lys Gly Ala Lys Asp Gly 
 65                  70                  75                  80 

Ala Leu Thr Leu Leu Leu Asp Glu Phe Glu Asn Met Ser Val Thr Arg 
                 85                  90                  95 

Ser Asn Ser Leu Arg Arg Asp Ser Pro Pro Pro Pro Ala Arg Ala Arg 
            100                 105                 110 

Gln Glu Asn Gly Met Pro Glu Glu Pro Ala Thr Thr Ala Arg Gly Gly 
        115                 120                 125 

Pro Gly Lys Ala Gly Ser Arg Gly Arg Phe Ala Gly His Ser Glu Ala 
    130                 135                 140 

Gly Gly Gly Ser Gly Asp Arg Arg Arg Ala Gly Pro Glu Lys Arg Pro 
145                 150                 155                 160 

Lys Ser Ser Arg Glu Gly Ser Gly Gly Pro Gln Glu Ser Ser Arg Asp 
                165                 170                 175 

Lys Arg Pro Leu Ser Gly Pro Asp Val Gly Thr Pro Gln Pro Ala Gly 
            180                 185                 190 

Leu Ala Ser Gly Ala Lys Leu Ala Ala Gly Arg Pro Phe Asn Thr Tyr 
        195                 200                 205 

Pro Arg Ala Asp Thr Asp His Pro Ser Arg Gly Ala Gln Gly Glu Pro 
    210                 215                 220 

His Asp Val Ala Pro Asn Gly Pro Ser Ala Gly Gly Leu Ala Ile Pro 
225                 230                 235                 240 

Gln Ser Ser Ser Ser Ser Ser Arg Pro Pro Thr Arg Ala Arg Gly Ala 
                245                 250                 255 

Pro Ser Pro Gly Val Leu Gly Pro His Ala Ser Glu Pro Gln Leu Ala 
            260                 265                 270 

Pro Pro Ala Cys Thr Pro Ala Ala Pro Ala Val Pro Gly Pro Pro Gly 
        275                 280                 285 

Pro Arg Ser Pro Gln Arg Glu Pro Gln Arg Val Ser His Glu Gln Phe 
    290                 295                 300 

Arg Ala Ala Leu Gln Leu Val Val Asp Pro Gly Asp Pro Arg Ser Tyr 
305                 310                 315                 320 

Leu Asp Asn Phe Ile Lys Ile Gly Glu Gly Ser Thr Gly Ile Val Cys 
                325                 330                 335 

Ile Ala Thr Val Arg Ser Ser Gly Lys Leu Val Ala Val Lys Lys Met 
            340                 345                 350 

Asp Leu Arg Lys Gln Gln Arg Arg Glu Leu Leu Phe Asn Glu Val Val 
        355                 360                 365 

Ile Met Arg Asp Tyr Gln His Glu Asn Val Val Glu Met Tyr Asn Ser 
    370                 375                 380 

Tyr Leu Val Gly Asp Glu Leu Trp Val Val Met Glu Phe Leu Glu Gly 
385                 390                 395                 400 

Gly Ala Leu Thr Asp Ile Val Thr His Thr Arg Met Asn Glu Glu Gln 
                405                 410                 415 

Ile Ala Ala Val Cys Leu Ala Val Leu Gln Ala Leu Ser Val Leu His 
            420                 425                 430 

Ala Gln Gly Val Ile His Arg Asp Ile Lys Ser Asp Ser Ile Leu Leu 
        435                 440                 445 

Thr His Asp Gly Arg Val Lys Leu Ser Asp Phe Gly Phe Cys Ala Gln 
    450                 455                 460 

Val Ser Lys Glu Val Pro Arg Arg Lys Ser Leu Val Gly Thr Pro Tyr 
465                 470                 475                 480 

Trp Met Ala Pro Glu Leu Ile Ser Arg Leu Pro Tyr Gly Pro Glu Val 
                485                 490                 495 

Asp Ile Trp Ser Leu Gly Ile Met Val Ile Glu Met Val Asp Gly Glu 
            500                 505                 510 

Pro Pro Tyr Phe Asn Glu Pro Pro Leu Lys Ala Met Lys Met Ile Arg 
        515                 520                 525 

Asp Asn Leu Pro Pro Arg Leu Lys Asn Leu His Lys Val Ser Pro Ser 
    530                 535                 540 

Leu Lys Gly Phe Leu Asp Arg Leu Leu Val Arg Asp Pro Ala Gln Arg 
545                 550                 555                 560 

Ala Thr Ala Ala Glu Leu Leu Lys His Pro Phe Leu Ala Lys Ala Gly 
                565                 570                 575 

Pro Pro Ala Ser Ile Val Pro Leu Met Arg Gln Asn 
            580                 585 

 
           
             2  
             635  
             PRT  
             Drosophila melanogaster  
           
            2 

Met Phe Ser Lys Lys Lys Lys Lys Pro Leu Ile Ser Met Pro Ser Asn 
  1               5                  10                  15 

Phe Glu His Arg Val His Thr Gly Phe Asp Lys Arg Glu Asn Lys Tyr 
             20                  25                  30 

Val Gly Leu Pro Leu Gln Trp Ala Ser Ile Val Gly Asn Asn Gln Ile 
         35                  40                  45 

Leu Lys Ser Ser Asn Arg Pro Leu Pro Leu Val Asp Pro Ser Glu Ile 
     50                  55                  60 

Thr Pro Thr Glu Ile Leu Asp Leu Lys Thr Ile Val Arg Pro His His 
 65                  70                  75                  80 

Asn Asn Asn Lys Ala Asp Thr Thr Ser Leu Asn Ser Ser Ser Thr Met 
                 85                  90                  95 

Met Met Gly Ser Met Ala Pro Met Asn Pro Met Ala Pro Gly Ala His 
            100                 105                 110 

Pro Met Met Ser His Gly Pro Gly Met Met Met Pro Pro Glu Thr Gly 
        115                 120                 125 

Gly Ile Val Leu Pro Lys Thr Ser His Val Ala Arg Ser Asn Ser Leu 
    130                 135                 140 

Arg Ser Ser Ser Pro Pro Arg Val Arg Arg Val Ala Asn Val Pro Pro 
145                 150                 155                 160 

Ser Val Pro Glu Glu Glu Gly Pro Pro Ala Ala Gly Thr Pro Gly Val 
                165                 170                 175 

Gly Gly Ala Ser Ser Gly Gly Phe Lys Pro Pro Gly Ala His Pro Ser 
            180                 185                 190 

Leu Leu Tyr Asn Ser Gln His Ala His Ala Asn Gly Ala Thr Gly Pro 
        195                 200                 205 

Leu Ala Val Arg Thr Asp Gln Thr Asn Leu Gln Gln Tyr Arg Ser Asn 
    210                 215                 220 

Leu Ala Pro Pro Ser Gly Gly Ser Met Pro Gln Gln Gln Gln Thr Ser 
225                 230                 235                 240 

Pro Val Gly Ser Val Ala Ser Gly Thr Arg Ser Asn His Ser His Thr 
                245                 250                 255 

Asn Asn Gly Asn Ser Gly Gly Ser Tyr Pro Pro Met Tyr Pro Thr Ser 
            260                 265                 270 

His Gln Gln Gln Gln Gln Gln Gln Gln Gln Ala Lys Gln Gly Gly Asp 
        275                 280                 285 

Gln Asn Gln Asn Pro Leu His Pro His Ala His Pro His Pro His His 
    290                 295                 300 

His Gln His Leu Ala Lys Ser Ala Ser Arg Ala Ser Ser Ser Ser Gly 
305                 310                 315                 320 

Gly Ala Ser Ser Ala Ala Gln Gln Ala Ser Gly Ala Ser Gly Gly Ala 
                325                 330                 335 

Ala Gly Gln Pro Lys Gln Asp Gln Arg Leu Thr His Glu Gln Phe Arg 
            340                 345                 350 

Ala Ala Leu Gln Met Val Val Ser Ala Gly Asp Pro Arg Glu Asn Leu 
        355                 360                 365 

Asp His Phe Asn Lys Ile Gly Glu Gly Ser Thr Gly Thr Val Cys Ile 
    370                 375                 380 

Ala Thr Asp Lys Ser Thr Gly Arg Gln Val Ala Val Lys Lys Met Asp 
385                 390                 395                 400 

Leu Arg Lys Gln Gln Arg Arg Glu Leu Leu Phe Asn Glu Val Val Ile 
                405                 410                 415 

Met Arg Asp Tyr His His Pro Asn Ile Val Glu Thr Tyr Ser Ser Phe 
            420                 425                 430 

Leu Val Asn Asp Glu Leu Trp Val Val Met Glu Tyr Leu Glu Gly Gly 
        435                 440                 445 

Ala Leu Thr Asp Ile Val Thr His Ser Arg Met Asp Glu Glu Gln Ile 
    450                 455                 460 

Ala Thr Val Cys Lys Gln Cys Leu Lys Ala Leu Ala Tyr Leu His Ser 
465                 470                 475                 480 

Gln Gly Val Ile His Arg Asp Ile Lys Ser Asp Ser Ile Leu Leu Ala 
                485                 490                 495 

Ala Asp Gly Arg Val Lys Leu Ser Asp Phe Gly Phe Cys Ala Gln Val 
            500                 505                 510 

Ser Gln Glu Leu Pro Lys Arg Lys Ser Leu Val Gly Thr Pro Tyr Trp 
        515                 520                 525 

Met Ser Pro Glu Val Ile Ser Arg Leu Pro Tyr Gly Pro Glu Val Asp 
    530                 535                 540 

Ile Trp Ser Leu Gly Ile Met Val Ile Glu Met Val Asp Gly Glu Pro 
545                 550                 555                 560 

Pro Phe Phe Asn Glu Pro Pro Leu Gln Ala Met Arg Arg Ile Arg Asp 
                565                 570                 575 

Met Gln Pro Pro Asn Leu Lys Asn Ala His Lys Val Ser Pro Arg Leu 
            580                 585                 590 

Gln Ser Phe Phe Asp Arg Met Leu Val Arg Asp Pro Ala Gln Arg Ala 
        595                 600                 605 

Thr Ala Ala Glu Leu Leu Ala His Pro Phe Leu Arg Gln Ala Gly Pro 
    610                 615                 620 

Pro Ser Leu Leu Val Pro Leu Met Arg Asn Ala 
625                 630                 635 

 
           
             3  
             2838  
             DNA  
             Homo sapiens  
             
               CDS  
               (211)..(1986)  
             
           
            3 

ccagtgtggt ggaattcgtc cgcggtggtg gcggtgcaag agagctgagg gaggcgcgag     60 

ggcgcggagt tccaggtcga gcagttaggc cgcgagcgac tgcggcgccg agccgatgag    120 

taacccgaag cccctagagg agtggtcacc tgcctgaggg cacttctgtc ccaccagcat    180 

cagaccaggc cgcaccgagt ccccggcacc atg ttt ggg aag agg aag aag cgg     234 
                                 Met Phe Gly Lys Arg Lys Lys Arg 
                                   1               5 

gtg gag atc tcc gcg ccg tcc aac ttc gag cac cgc gtg cac acg ggc      282 
Val Glu Ile Ser Ala Pro Ser Asn Phe Glu His Arg Val His Thr Gly 
     10                  15                  20 

ttc gac cag cac gag cag aag ttc acg ggg ctg ccc cgc cag tgg cag      330 
Phe Asp Gln His Glu Gln Lys Phe Thr Gly Leu Pro Arg Gln Trp Gln 
 25                  30                  35                  40 

agc ctg atc gag gag tcg gct cgc cgg ccc aag ccc ctc gtc gac ccc      378 
Ser Leu Ile Glu Glu Ser Ala Arg Arg Pro Lys Pro Leu Val Asp Pro 
                 45                  50                  55 

gcc tgc atc acc tcc atc cag ccc ggg gcc ccc aag acc atc gtg cgg      426 
Ala Cys Ile Thr Ser Ile Gln Pro Gly Ala Pro Lys Thr Ile Val Arg 
             60                  65                  70 

ggc agc aaa ggt gcc aaa gat ggg gcc ctc acg ctg ctg ctg gac gag      474 
Gly Ser Lys Gly Ala Lys Asp Gly Ala Leu Thr Leu Leu Leu Asp Glu 
         75                  80                  85 

ttt gag aac atg tcg gtg aca cgc tcc aac tcc ctg cgg aga gac agc      522 
Phe Glu Asn Met Ser Val Thr Arg Ser Asn Ser Leu Arg Arg Asp Ser 
     90                  95                 100 

ccg ccg ccg ccc gcc cgt gcc cgc cag gaa aat ggg atg cca gag gag      570 
Pro Pro Pro Pro Ala Arg Ala Arg Gln Glu Asn Gly Met Pro Glu Glu 
105                 110                 115                 120 

ccg gcc acc acg gcc aga ggg ggc cca ggg aag gca ggc agc cga ggc      618 
Pro Ala Thr Thr Ala Arg Gly Gly Pro Gly Lys Ala Gly Ser Arg Gly 
                125                 130                 135 

cgg ttc gcc ggt cac agc gag gcg ggt ggc ggc agt ggt gac agg cga      666 
Arg Phe Ala Gly His Ser Glu Ala Gly Gly Gly Ser Gly Asp Arg Arg 
            140                 145                 150 

cgg gcg ggg cca gag aag agg ccc aag tct tcc agg gag ggc tca ggg      714 
Arg Ala Gly Pro Glu Lys Arg Pro Lys Ser Ser Arg Glu Gly Ser Gly 
        155                 160                 165 

ggt ccc cag gag tcc tcc cgg gac aaa cgc ccc ctc tcc ggg cct gat      762 
Gly Pro Gln Glu Ser Ser Arg Asp Lys Arg Pro Leu Ser Gly Pro Asp 
    170                 175                 180 

gtc ggc acc ccc cag cct gct ggt ctg gcc agt ggg gcg aaa ctg gca      810 
Val Gly Thr Pro Gln Pro Ala Gly Leu Ala Ser Gly Ala Lys Leu Ala 
185                 190                 195                 200 

gct ggc cgg ccc ttt aac acc tac ccg agg gct gac acg gac cac cca      858 
Ala Gly Arg Pro Phe Asn Thr Tyr Pro Arg Ala Asp Thr Asp His Pro 
                205                 210                 215 

tcc cgg ggt gcc cag ggg gag cct cat gac gtg gcc cct aac ggg cca      906 
Ser Arg Gly Ala Gln Gly Glu Pro His Asp Val Ala Pro Asn Gly Pro 
            220                 225                 230 

tca gcg ggg ggc ctg gcc atc ccc cag tcc tcc tcc tcc tcc tcc cgg      954 
Ser Ala Gly Gly Leu Ala Ile Pro Gln Ser Ser Ser Ser Ser Ser Arg 
        235                 240                 245 

cct ccc acc cga gcc cga ggt gcc ccc agc cct gga gtg ctg gga ccc     1002 
Pro Pro Thr Arg Ala Arg Gly Ala Pro Ser Pro Gly Val Leu Gly Pro 
    250                 255                 260 

cac gcc tca gag ccc cag ctg gcc cct cca gcc tgc acc ccc gcc gcc     1050 
His Ala Ser Glu Pro Gln Leu Ala Pro Pro Ala Cys Thr Pro Ala Ala 
265                 270                 275                 280 

cct gct gtt cct ggg ccc cct ggc ccc cgc tca cca cag cgg gag cca     1098 
Pro Ala Val Pro Gly Pro Pro Gly Pro Arg Ser Pro Gln Arg Glu Pro 
                285                 290                 295 

cag cga gta tcc cat gag cag ttc cgg gct gcc ctg cag ctg gtg gtg     1146 
Gln Arg Val Ser His Glu Gln Phe Arg Ala Ala Leu Gln Leu Val Val 
            300                 305                 310 

gac cca ggc gac ccc cgc tcc tac ctg gac aac ttc atc aag att ggc     1194 
Asp Pro Gly Asp Pro Arg Ser Tyr Leu Asp Asn Phe Ile Lys Ile Gly 
        315                 320                 325 

gag ggc tcc acg ggc atc gtg tgc atc gcc acc gtg cgc agc tcg ggc     1242 
Glu Gly Ser Thr Gly Ile Val Cys Ile Ala Thr Val Arg Ser Ser Gly 
    330                 335                 340 

aag ctg gtg gcc gtc aag aag atg gac ctg cgc aag cag cag agg cgc     1290 
Lys Leu Val Ala Val Lys Lys Met Asp Leu Arg Lys Gln Gln Arg Arg 
345                 350                 355                 360 

gag ctg ctc ttc aac gag gtg gta atc atg agg gac tac cag cac gag     1338 
Glu Leu Leu Phe Asn Glu Val Val Ile Met Arg Asp Tyr Gln His Glu 
                365                 370                 375 

aat gtg gtg gag atg tac aac agc tac ctg gtg ggg gac gag ctc tgg     1386 
Asn Val Val Glu Met Tyr Asn Ser Tyr Leu Val Gly Asp Glu Leu Trp 
            380                 385                 390 

gtg gtc atg gag ttc ctg gaa gga ggc gcc ctc acc gac atc gtc acc     1434 
Val Val Met Glu Phe Leu Glu Gly Gly Ala Leu Thr Asp Ile Val Thr 
        395                 400                 405 

cac acc agg atg aac gag gag cag atc gcg gcc gtg tgc ctt gca gtg     1482 
His Thr Arg Met Asn Glu Glu Gln Ile Ala Ala Val Cys Leu Ala Val 
    410                 415                 420 

ctg cag gcc ctg tcg gtg ctc cac gcc cag ggc gtc atc cac cgg gac     1530 
Leu Gln Ala Leu Ser Val Leu His Ala Gln Gly Val Ile His Arg Asp 
425                 430                 435                 440 

atc aag agc gac tcg atc ctg ctg acc cat gat ggc agg gtg aag ctg     1578 
Ile Lys Ser Asp Ser Ile Leu Leu Thr His Asp Gly Arg Val Lys Leu 
                445                 450                 455 

tca gac ttt ggg ttc tgc gcc cag gtg agc aag gaa gtg ccc cga agg     1626 
Ser Asp Phe Gly Phe Cys Ala Gln Val Ser Lys Glu Val Pro Arg Arg 
            460                 465                 470 

aag tcg ctg gtc ggc acg ccc tac tgg atg gcc cca gag ctc atc tcc     1674 
Lys Ser Leu Val Gly Thr Pro Tyr Trp Met Ala Pro Glu Leu Ile Ser 
        475                 480                 485 

cgc ctt ccc tac ggg cca gag gta gac atc tgg tcg ctg ggg ata atg     1722 
Arg Leu Pro Tyr Gly Pro Glu Val Asp Ile Trp Ser Leu Gly Ile Met 
    490                 495                 500 

gtg att gag atg gtg gac gga gag ccc ccc tac ttc aac gag cca ccc     1770 
Val Ile Glu Met Val Asp Gly Glu Pro Pro Tyr Phe Asn Glu Pro Pro 
505                 510                 515                 520 

ctc aaa gcc atg aag atg att cgg gac aac ctg cca ccc cga ctg aag     1818 
Leu Lys Ala Met Lys Met Ile Arg Asp Asn Leu Pro Pro Arg Leu Lys 
                525                 530                 535 

aac ctg cac aag gtg tcg cca tcc ctg aag ggc ttc ctg gac cgc ctg     1866 
Asn Leu His Lys Val Ser Pro Ser Leu Lys Gly Phe Leu Asp Arg Leu 
            540                 545                 550 

ctg gtg cga gac cct gcc cag cgg gcc acg gca gcc gag ctg ctg aag     1914 
Leu Val Arg Asp Pro Ala Gln Arg Ala Thr Ala Ala Glu Leu Leu Lys 
        555                 560                 565 

cac cca ttc ctg gcc aag gca ggg ccg cct gcc agc atc gtg ccc ctc     1962 
His Pro Phe Leu Ala Lys Ala Gly Pro Pro Ala Ser Ile Val Pro Leu 
    570                 575                 580 

atg cgc cag aac cgc acc aga tga ggcccagcgc ccttcccctc aaccaaagag    2016 
Met Arg Gln Asn Arg Thr Arg 
585                 590 

ccccccgggt cacccccgcc ccactgaggc cagtaggggg ccaggcctcc cactcctccc   2076 

agcccgggag atgctccgcg tggcaccacc ctccttgctg ggggtagatg agaccctact   2136 

actgaactcc agttttgatc tcgtgacttt tagaaaaaca cagggactcg tgggagcaag   2196 

cgaggctccc aggaccccca ccctctggga caggccctcc cccatgttct tctgtctcca   2256 

ggaagggcag cggccctccc atcactggaa gtctgcagtg ggggtcgctg ggggtggaga   2316 

gaacactaag aggtgaacat gtatgagtgt gtgcacgcgt gtgagtgtgc atgtgtgtgt   2376 

gtgtgcaaag gtccagccac cccgtcctcc agcccgcaag gggtgtctgg cgccttgcct   2436 

gacacccagc cccctctccc cctgagccat tgtgggggtc gatcatgaat gtccgaagag   2496 

tggccttttc ccgtagccct gcgccccctt tctgtggctg gatggggaga caggtcaggg   2556 

ccccccaccc tctccagccc ctgcagcaaa tgactactgc acctggacag cctcctcttt   2616 

tctagaagtc tatttatatt gtcattttat aacactctag cccctgccct tattggggga   2676 

cagatggtcc ctgtcctgcg gggtggccct ggcagaacca ctgcctgaag aaccaggttc   2736 

ctgcccggtc agcgcagccc cagcccgccc acccctgcct cgagttagtt ttacaattaa   2796 

aacattgtct tgtttaaaaa aaaaaaaaaa aaaaaaaaaa aa                      2838 

 
           
             4  
             591  
             PRT  
             Homo sapiens  
           
            4 

Met Phe Gly Lys Arg Lys Lys Arg Val Glu Ile Ser Ala Pro Ser Asn 
  1               5                  10                  15 

Phe Glu His Arg Val His Thr Gly Phe Asp Gln His Glu Gln Lys Phe 
             20                  25                  30 

Thr Gly Leu Pro Arg Gln Trp Gln Ser Leu Ile Glu Glu Ser Ala Arg 
         35                  40                  45 

Arg Pro Lys Pro Leu Val Asp Pro Ala Cys Ile Thr Ser Ile Gln Pro 
     50                  55                  60 

Gly Ala Pro Lys Thr Ile Val Arg Gly Ser Lys Gly Ala Lys Asp Gly 
 65                  70                  75                  80 

Ala Leu Thr Leu Leu Leu Asp Glu Phe Glu Asn Met Ser Val Thr Arg 
                 85                  90                  95 

Ser Asn Ser Leu Arg Arg Asp Ser Pro Pro Pro Pro Ala Arg Ala Arg 
            100                 105                 110 

Gln Glu Asn Gly Met Pro Glu Glu Pro Ala Thr Thr Ala Arg Gly Gly 
        115                 120                 125 

Pro Gly Lys Ala Gly Ser Arg Gly Arg Phe Ala Gly His Ser Glu Ala 
    130                 135                 140 

Gly Gly Gly Ser Gly Asp Arg Arg Arg Ala Gly Pro Glu Lys Arg Pro 
145                 150                 155                 160 

Lys Ser Ser Arg Glu Gly Ser Gly Gly Pro Gln Glu Ser Ser Arg Asp 
                165                 170                 175 

Lys Arg Pro Leu Ser Gly Pro Asp Val Gly Thr Pro Gln Pro Ala Gly 
            180                 185                 190 

Leu Ala Ser Gly Ala Lys Leu Ala Ala Gly Arg Pro Phe Asn Thr Tyr 
        195                 200                 205 

Pro Arg Ala Asp Thr Asp His Pro Ser Arg Gly Ala Gln Gly Glu Pro 
    210                 215                 220 

His Asp Val Ala Pro Asn Gly Pro Ser Ala Gly Gly Leu Ala Ile Pro 
225                 230                 235                 240 

Gln Ser Ser Ser Ser Ser Ser Arg Pro Pro Thr Arg Ala Arg Gly Ala 
                245                 250                 255 

Pro Ser Pro Gly Val Leu Gly Pro His Ala Ser Glu Pro Gln Leu Ala 
            260                 265                 270 

Pro Pro Ala Cys Thr Pro Ala Ala Pro Ala Val Pro Gly Pro Pro Gly 
        275                 280                 285 

Pro Arg Ser Pro Gln Arg Glu Pro Gln Arg Val Ser His Glu Gln Phe 
    290                 295                 300 

Arg Ala Ala Leu Gln Leu Val Val Asp Pro Gly Asp Pro Arg Ser Tyr 
305                 310                 315                 320 

Leu Asp Asn Phe Ile Lys Ile Gly Glu Gly Ser Thr Gly Ile Val Cys 
                325                 330                 335 

Ile Ala Thr Val Arg Ser Ser Gly Lys Leu Val Ala Val Lys Lys Met 
            340                 345                 350 

Asp Leu Arg Lys Gln Gln Arg Arg Glu Leu Leu Phe Asn Glu Val Val 
        355                 360                 365 

Ile Met Arg Asp Tyr Gln His Glu Asn Val Val Glu Met Tyr Asn Ser 
    370                 375                 380 

Tyr Leu Val Gly Asp Glu Leu Trp Val Val Met Glu Phe Leu Glu Gly 
385                 390                 395                 400 

Gly Ala Leu Thr Asp Ile Val Thr His Thr Arg Met Asn Glu Glu Gln 
                405                 410                 415 

Ile Ala Ala Val Cys Leu Ala Val Leu Gln Ala Leu Ser Val Leu His 
            420                 425                 430 

Ala Gln Gly Val Ile His Arg Asp Ile Lys Ser Asp Ser Ile Leu Leu 
        435                 440                 445 

Thr His Asp Gly Arg Val Lys Leu Ser Asp Phe Gly Phe Cys Ala Gln 
    450                 455                 460 

Val Ser Lys Glu Val Pro Arg Arg Lys Ser Leu Val Gly Thr Pro Tyr 
465                 470                 475                 480 

Trp Met Ala Pro Glu Leu Ile Ser Arg Leu Pro Tyr Gly Pro Glu Val 
                485                 490                 495 

Asp Ile Trp Ser Leu Gly Ile Met Val Ile Glu Met Val Asp Gly Glu 
            500                 505                 510 

Pro Pro Tyr Phe Asn Glu Pro Pro Leu Lys Ala Met Lys Met Ile Arg 
        515                 520                 525 

Asp Asn Leu Pro Pro Arg Leu Lys Asn Leu His Lys Val Ser Pro Ser 
    530                 535                 540 

Leu Lys Gly Phe Leu Asp Arg Leu Leu Val Arg Asp Pro Ala Gln Arg 
545                 550                 555                 560 

Ala Thr Ala Ala Glu Leu Leu Lys His Pro Phe Leu Ala Lys Ala Gly 
                565                 570                 575 

Pro Pro Ala Ser Ile Val Pro Leu Met Arg Gln Asn Arg Thr Arg 
            580                 585                 590 

 
           
             5  
             3351  
             DNA  
             Drosophila melanogaster  
             
               CDS  
               (493)..(2412)  
             
           
            5 

gcacgagcac cacctctatt tctggcatcg gcaattattt ccctaggttt attatttttt     60 

tgttaagcga attgcttgtg cgctcgtgct gtcgccataa tgtgtgcaat gtgtgccagc    120 

cagaggagca agtggtgaag gtgtatcgct tgtgcgcatc tcgatttgtc catccatatc    180 

caattgaatc ccagtttccc agtcgcgtcc gctgctatgc gtgtgtgtgt gcgtgcgtgc    240 

ccgtgtccct gcatgtgtgt gtgtgagtga gatcgggaag aagttgtgtg ctgttgttgt    300 

tgttgtttct gtcggatttg ccgttgccat cgtttgttgt ttgtgcattt tccattgtcg    360 

ggcgacgcaa caaaaagaaa aaattacaca ttaaaaatag agagcaagag cgagagggaa    420 

ggagtggtgt ggagtggaaa agggcggtaa aagggcagaa atttggagca cgggaaatcc    480 

gagtcgctga ag atg ttc tcg aag aag aaa aag aaa ccg ctg atc tcg atg    531 
              Met Phe Ser Lys Lys Lys Lys Lys Pro Leu Ile Ser Met 
                1               5                  10 

ccc agc aat ttt gag cat cgt gtg cac acg ggc ttc gac aag cgg gag      579 
Pro Ser Asn Phe Glu His Arg Val His Thr Gly Phe Asp Lys Arg Glu 
     15                  20                  25 

aac aaa tat gtt ggc ctg ccc ctc caa tgg gcg tcc att gtg ggc aac      627 
Asn Lys Tyr Val Gly Leu Pro Leu Gln Trp Ala Ser Ile Val Gly Asn 
 30                  35                  40                  45 

aat cag ata ctc aag tcc tcc aac cgc ccg ctg cca ctg gtc gat ccc      675 
Asn Gln Ile Leu Lys Ser Ser Asn Arg Pro Leu Pro Leu Val Asp Pro 
                 50                  55                  60 

tcg gag att acg ccc acc gag att ctc gat ctg aag act att gtg cgt      723 
Ser Glu Ile Thr Pro Thr Glu Ile Leu Asp Leu Lys Thr Ile Val Arg 
             65                  70                  75 

ccg cat cac aac aac aac aag gcg gac acc acc tcg ctc aac agc agc      771 
Pro His His Asn Asn Asn Lys Ala Asp Thr Thr Ser Leu Asn Ser Ser 
         80                  85                  90 

agc aca atg atg atg ggc tca atg gcg ccg atg aat ccc atg gca cct      819 
Ser Thr Met Met Met Gly Ser Met Ala Pro Met Asn Pro Met Ala Pro 
     95                 100                 105 

ggc gca cac cca atg atg agc cat ggc ccc gga atg atg atg cca ccc      867 
Gly Ala His Pro Met Met Ser His Gly Pro Gly Met Met Met Pro Pro 
110                 115                 120                 125 

gag acg ggc ggc ata gtc ctg ccg aag acc tct cac gtg gcc aga tcc      915 
Glu Thr Gly Gly Ile Val Leu Pro Lys Thr Ser His Val Ala Arg Ser 
                130                 135                 140 

aat tcg ctg cgg agt tcc agt ccg ccg cga gtg cga cgg gta gcc aat      963 
Asn Ser Leu Arg Ser Ser Ser Pro Pro Arg Val Arg Arg Val Ala Asn 
            145                 150                 155 

gtg ccg cca tcg gtg ccg gag gag gag gga cca ccg gca gct gga aca     1011 
Val Pro Pro Ser Val Pro Glu Glu Glu Gly Pro Pro Ala Ala Gly Thr 
        160                 165                 170 

ccg gga gta ggt gga gct agt agc ggt ggc ttt aag cca ccc ggt gcc     1059 
Pro Gly Val Gly Gly Ala Ser Ser Gly Gly Phe Lys Pro Pro Gly Ala 
    175                 180                 185 

cat ccc tcc ctg ctc tat aac agt cag cat gcg cac gcg aat gga gca     1107 
His Pro Ser Leu Leu Tyr Asn Ser Gln His Ala His Ala Asn Gly Ala 
190                 195                 200                 205 

aca gga cca ctg gcc gtg cgc acg gat caa acc aac ctg cag cag tat     1155 
Thr Gly Pro Leu Ala Val Arg Thr Asp Gln Thr Asn Leu Gln Gln Tyr 
                210                 215                 220 

cgc agc aat ctg gcc ccg cca tcc ggc ggc tcc atg ccc cag caa cag     1203 
Arg Ser Asn Leu Ala Pro Pro Ser Gly Gly Ser Met Pro Gln Gln Gln 
            225                 230                 235 

cag act tcg ccc gtg ggt tcg gtg gcc agt ggc acg cga tcc aat cac     1251 
Gln Thr Ser Pro Val Gly Ser Val Ala Ser Gly Thr Arg Ser Asn His 
        240                 245                 250 

tcg cac acg aac aat ggc aac agc ggc ggc agc tat cct ccc atg tat     1299 
Ser His Thr Asn Asn Gly Asn Ser Gly Gly Ser Tyr Pro Pro Met Tyr 
    255                 260                 265 

ccc aca agc cat cag cag cag cag cag cag caa cag cag gcc aaa cag     1347 
Pro Thr Ser His Gln Gln Gln Gln Gln Gln Gln Gln Gln Ala Lys Gln 
270                 275                 280                 285 

ggt ggc gat cag aac caa aat cct ctg cat ccg cat gct cat ccg cat     1395 
Gly Gly Asp Gln Asn Gln Asn Pro Leu His Pro His Ala His Pro His 
                290                 295                 300 

ccg cac cat cac caa cat ttg gcc aag tcg gcg tcc agg gca tcc agt     1443 
Pro His His His Gln His Leu Ala Lys Ser Ala Ser Arg Ala Ser Ser 
            305                 310                 315 

tcc agc ggg gga gcg agc agt gct gcc cag cag gcg agc ggc gcc agt     1491 
Ser Ser Gly Gly Ala Ser Ser Ala Ala Gln Gln Ala Ser Gly Ala Ser 
        320                 325                 330 

gga gga gcg gcg ggt cag cca aag cag gac caa cga ctc acc cac gaa     1539 
Gly Gly Ala Ala Gly Gln Pro Lys Gln Asp Gln Arg Leu Thr His Glu 
    335                 340                 345 

cag ttc cgt gcc gcc ctt cag atg gtg gtc tcc gcc ggt gat cca cgc     1587 
Gln Phe Arg Ala Ala Leu Gln Met Val Val Ser Ala Gly Asp Pro Arg 
350                 355                 360                 365 

gag aac ctc gac cac ttt aac aaa ata ggc gag ggc tcc acg ggc acc     1635 
Glu Asn Leu Asp His Phe Asn Lys Ile Gly Glu Gly Ser Thr Gly Thr 
                370                 375                 380 

gta tgc att gcc acg gac aaa tcc aca ggt cgc cag gtg gcc gtg aag     1683 
Val Cys Ile Ala Thr Asp Lys Ser Thr Gly Arg Gln Val Ala Val Lys 
            385                 390                 395 

aag atg gat ctg cgc aaa cag cag cga cgg gag ctg cta ttc aac gag     1731 
Lys Met Asp Leu Arg Lys Gln Gln Arg Arg Glu Leu Leu Phe Asn Glu 
        400                 405                 410 

gtc gtc atc atg cgg gac tac cat cat ccc aat atc gtg gag aca tac     1779 
Val Val Ile Met Arg Asp Tyr His His Pro Asn Ile Val Glu Thr Tyr 
    415                 420                 425 

tcc agc ttt ctg gtc aac gat gag ctt tgg gtg gtg atg gag tac ctc     1827 
Ser Ser Phe Leu Val Asn Asp Glu Leu Trp Val Val Met Glu Tyr Leu 
430                 435                 440                 445 

gag ggc ggc gcc ctc acc gat att gtc acc cat tcg cgc atg gac gag     1875 
Glu Gly Gly Ala Leu Thr Asp Ile Val Thr His Ser Arg Met Asp Glu 
                450                 455                 460 

gag cag ata gcc acc gtc tgc aag caa tgc ttg aag gct ttg gcc tat     1923 
Glu Gln Ile Ala Thr Val Cys Lys Gln Cys Leu Lys Ala Leu Ala Tyr 
            465                 470                 475 

ttg cac tca cag ggc gtc att cat cgc gac atc aag tcg gac tcg att     1971 
Leu His Ser Gln Gly Val Ile His Arg Asp Ile Lys Ser Asp Ser Ile 
        480                 485                 490 

ctg ctg gcc gcc gat ggt cgc gtg aag cta tcg gac ttt gga ttc tgc     2019 
Leu Leu Ala Ala Asp Gly Arg Val Lys Leu Ser Asp Phe Gly Phe Cys 
    495                 500                 505 

gcc cag gtg tca cag gag ctg ccg aag cgc aag agt ctg gtt ggc acg     2067 
Ala Gln Val Ser Gln Glu Leu Pro Lys Arg Lys Ser Leu Val Gly Thr 
510                 515                 520                 525 

ccg tat tgg atg tcg ccg gag gtc ata tcg cgc ctg ccg tac ggc ccg     2115 
Pro Tyr Trp Met Ser Pro Glu Val Ile Ser Arg Leu Pro Tyr Gly Pro 
                530                 535                 540 

gaa gtg gat atc tgg tcg ctg ggc atc atg gtc atc gag atg gtg gac     2163 
Glu Val Asp Ile Trp Ser Leu Gly Ile Met Val Ile Glu Met Val Asp 
            545                 550                 555 

ggc gag ccg ccg ttc ttc aac gaa ccg ccg ctg cag gcg atg cgt cgc     2211 
Gly Glu Pro Pro Phe Phe Asn Glu Pro Pro Leu Gln Ala Met Arg Arg 
        560                 565                 570 

atc cgt gac atg cag ccg ccg aac ctg aag aac gcg cac aag gtc tcg     2259 
Ile Arg Asp Met Gln Pro Pro Asn Leu Lys Asn Ala His Lys Val Ser 
    575                 580                 585 

ccg cgc ctg cag tct ttc ttc gac cgg atg ctg gtg cgg gat ccg gcg     2307 
Pro Arg Leu Gln Ser Phe Phe Asp Arg Met Leu Val Arg Asp Pro Ala 
590                 595                 600                 605 

cag cgt gcc acc gcc gcc gag ctg ctg gcc cat ccc ttc ctg cgc cag     2355 
Gln Arg Ala Thr Ala Ala Glu Leu Leu Ala His Pro Phe Leu Arg Gln 
                610                 615                 620 

gca gga ccg ccg tcg ctg ctg gtt ccg ctg atg cgc aat gcg cga cac     2403 
Ala Gly Pro Pro Ser Leu Leu Val Pro Leu Met Arg Asn Ala Arg His 
            625                 630                 635 

cat ccg tag ttggatgcca ctggacaacc cagacgatcc ggctgctcga             2452 
His Pro 
    640 

tgaaagcaat cattcccaca gacaacggat cccggcccgt gggcatccat ctattaaatt   2512 

ctagtattaa cattcagttt ggcagctgta cttaaatccc cggagaatcc tcgaaacgtt   2572 

gtaaagtcat ttaagctccg cacatccata tccatatcca tgcccactaa cgttccagaa   2632 

gagaacgcaa tgtgcctttc acaagcctcg aaatcgaaat tgaaatcgca tcctgcaacc   2692 

gatacggatg gagaaacgat atagcattta tctgcccgag aaccacattt ttatatatat   2752 

atatatatat acatatatga gtatttcagc attttgctct ctgtctcttg tcaatattga   2812 

tttgtgtgcg tgtgtgtttg atttttttta accagcccat aaaacattgt tcgaatatat   2872 

acatatatac gctaagagct aatgttaaaa atctatatat atatatatat acacatatat   2932 

tttttataaa aacaaatcct aattctaagc aaaagcgaaa caacgaaggt gtgaatcgaa   2992 

gggagacaga aaacgaaaga gaaagagata gcgttaggcg gaagagcata tacaatttcg   3052 

acataaatat tttttaaaca ttttacattt tacattggtg tgaaaagttt gtacgctcca   3112 

gacatacaca tatgcatata tctaaacaca ttcccgattt cgaaaatatt ttagaacttt   3172 

tagcttacat tctttgttct gtttttttga tttaacccgt gtaattatta attaatgaat   3232 

aaacttaaat gcgaaaatgt taaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa   3292 

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaa    3351 

 
           
             6  
             639  
             PRT  
             Drosophila melanogaster  
           
            6 

Met Phe Ser Lys Lys Lys Lys Lys Pro Leu Ile Ser Met Pro Ser Asn 
  1               5                  10                  15 

Phe Glu His Arg Val His Thr Gly Phe Asp Lys Arg Glu Asn Lys Tyr 
             20                  25                  30 

Val Gly Leu Pro Leu Gln Trp Ala Ser Ile Val Gly Asn Asn Gln Ile 
         35                  40                  45 

Leu Lys Ser Ser Asn Arg Pro Leu Pro Leu Val Asp Pro Ser Glu Ile 
     50                  55                  60 

Thr Pro Thr Glu Ile Leu Asp Leu Lys Thr Ile Val Arg Pro His His 
 65                  70                  75                  80 

Asn Asn Asn Lys Ala Asp Thr Thr Ser Leu Asn Ser Ser Ser Thr Met 
                 85                  90                  95 

Met Met Gly Ser Met Ala Pro Met Asn Pro Met Ala Pro Gly Ala His 
            100                 105                 110 

Pro Met Met Ser His Gly Pro Gly Met Met Met Pro Pro Glu Thr Gly 
        115                 120                 125 

Gly Ile Val Leu Pro Lys Thr Ser His Val Ala Arg Ser Asn Ser Leu 
    130                 135                 140 

Arg Ser Ser Ser Pro Pro Arg Val Arg Arg Val Ala Asn Val Pro Pro 
145                 150                 155                 160 

Ser Val Pro Glu Glu Glu Gly Pro Pro Ala Ala Gly Thr Pro Gly Val 
                165                 170                 175 

Gly Gly Ala Ser Ser Gly Gly Phe Lys Pro Pro Gly Ala His Pro Ser 
            180                 185                 190 

Leu Leu Tyr Asn Ser Gln His Ala His Ala Asn Gly Ala Thr Gly Pro 
        195                 200                 205 

Leu Ala Val Arg Thr Asp Gln Thr Asn Leu Gln Gln Tyr Arg Ser Asn 
    210                 215                 220 

Leu Ala Pro Pro Ser Gly Gly Ser Met Pro Gln Gln Gln Gln Thr Ser 
225                 230                 235                 240 

Pro Val Gly Ser Val Ala Ser Gly Thr Arg Ser Asn His Ser His Thr 
                245                 250                 255 

Asn Asn Gly Asn Ser Gly Gly Ser Tyr Pro Pro Met Tyr Pro Thr Ser 
            260                 265                 270 

His Gln Gln Gln Gln Gln Gln Gln Gln Gln Ala Lys Gln Gly Gly Asp 
        275                 280                 285 

Gln Asn Gln Asn Pro Leu His Pro His Ala His Pro His Pro His His 
    290                 295                 300 

His Gln His Leu Ala Lys Ser Ala Ser Arg Ala Ser Ser Ser Ser Gly 
305                 310                 315                 320 

Gly Ala Ser Ser Ala Ala Gln Gln Ala Ser Gly Ala Ser Gly Gly Ala 
                325                 330                 335 

Ala Gly Gln Pro Lys Gln Asp Gln Arg Leu Thr His Glu Gln Phe Arg 
            340                 345                 350 

Ala Ala Leu Gln Met Val Val Ser Ala Gly Asp Pro Arg Glu Asn Leu 
        355                 360                 365 

Asp His Phe Asn Lys Ile Gly Glu Gly Ser Thr Gly Thr Val Cys Ile 
    370                 375                 380 

Ala Thr Asp Lys Ser Thr Gly Arg Gln Val Ala Val Lys Lys Met Asp 
385                 390                 395                 400 

Leu Arg Lys Gln Gln Arg Arg Glu Leu Leu Phe Asn Glu Val Val Ile 
                405                 410                 415 

Met Arg Asp Tyr His His Pro Asn Ile Val Glu Thr Tyr Ser Ser Phe 
            420                 425                 430 

Leu Val Asn Asp Glu Leu Trp Val Val Met Glu Tyr Leu Glu Gly Gly 
        435                 440                 445 

Ala Leu Thr Asp Ile Val Thr His Ser Arg Met Asp Glu Glu Gln Ile 
    450                 455                 460 

Ala Thr Val Cys Lys Gln Cys Leu Lys Ala Leu Ala Tyr Leu His Ser 
465                 470                 475                 480 

Gln Gly Val Ile His Arg Asp Ile Lys Ser Asp Ser Ile Leu Leu Ala 
                485                 490                 495 

Ala Asp Gly Arg Val Lys Leu Ser Asp Phe Gly Phe Cys Ala Gln Val 
            500                 505                 510 

Ser Gln Glu Leu Pro Lys Arg Lys Ser Leu Val Gly Thr Pro Tyr Trp 
        515                 520                 525 

Met Ser Pro Glu Val Ile Ser Arg Leu Pro Tyr Gly Pro Glu Val Asp 
    530                 535                 540 

Ile Trp Ser Leu Gly Ile Met Val Ile Glu Met Val Asp Gly Glu Pro 
545                 550                 555                 560 

Pro Phe Phe Asn Glu Pro Pro Leu Gln Ala Met Arg Arg Ile Arg Asp 
                565                 570                 575 

Met Gln Pro Pro Asn Leu Lys Asn Ala His Lys Val Ser Pro Arg Leu 
            580                 585                 590 

Gln Ser Phe Phe Asp Arg Met Leu Val Arg Asp Pro Ala Gln Arg Ala 
        595                 600                 605 

Thr Ala Ala Glu Leu Leu Ala His Pro Phe Leu Arg Gln Ala Gly Pro 
    610                 615                 620 

Pro Ser Leu Leu Val Pro Leu Met Arg Asn Ala Arg His His Pro 
625                 630                 635 

 
           
             7  
             5790  
             DNA  
             Homo sapiens  
             
               CDS  
               (746)..(5314)  
             
           
            7 

aattggctca tttaagaatt tcaaaacatt taatgtaaaa gctttttttt ttttaaggaa     60 

gtccataaat tttggttccc agggttgcac tggacttgga aggagtgctg ttgtgtacat    120 

actattgtat ggttttattt attattttac tgtacaaatc agccgaaaga atttttccaa    180 

gtgccatttc ggatttatta atcctttttt ttttcctttc ctcaaagata tttgctgttg    240 

tcatattaag cattggagac tagaaaatta ctttccccct ttgagctaga gggtctcttg    300 

ccaacagaag gacagctgag aaagctggat ttaaaggatg gttttatctg tactttgcag    360 

ttaacagtga tattttgaag gcacattttt ctgtgattca tttttttttg gccatagtgc    420 

taaccttgaa gagattcgtg gctgggtttt tggtttctga gaaggtcgta gtttttcctc    480 

ttttcctttt tttttttctt ttttcttttc ttttcttttt ttttaaagcg ggggagggga    540 

agaggggctg agaaaggaaa tcatgttcac tggtagaagt agagtggagc atcagttacc    600 

agggtcctga gagctggagg agaaaggatt ctatcttcaa gttgggaggc cctcctctca    660 

ccttgctcaa aaattgcaag cgattcaatc ctgatcaaga caccaaagct acaggattct    720 

ggaaccgtgg agacaccgag aaacc atg agt gta agg tta ccc cag agt ata      772 
                            Met Ser Val Arg Leu Pro Gln Ser Ile 
                              1               5 

gac agg tta agt agc ctg tct tct ctg gga gat tct gca cca gag cgc      820 
Asp Arg Leu Ser Ser Leu Ser Ser Leu Gly Asp Ser Ala Pro Glu Arg 
 10                  15                  20                  25 

aag tcc cct tcc cac cat cgc cag cct tcg gat gcc tct gag aca aca      868 
Lys Ser Pro Ser His His Arg Gln Pro Ser Asp Ala Ser Glu Thr Thr 
                 30                  35                  40 

ggt ctc gtt caa cgc tgt gtc att atc caa aag gac cag cat ggc ttc      916 
Gly Leu Val Gln Arg Cys Val Ile Ile Gln Lys Asp Gln His Gly Phe 
             45                  50                  55 

ggc ttc aca gtc agt ggg gat cgc att gtt ctg gtg cag tct gtg cgg      964 
Gly Phe Thr Val Ser Gly Asp Arg Ile Val Leu Val Gln Ser Val Arg 
         60                  65                  70 

cct gga ggt gca gcc atg aag gcc ggt gtg aaa gag ggc gac cgg atc     1012 
Pro Gly Gly Ala Ala Met Lys Ala Gly Val Lys Glu Gly Asp Arg Ile 
     75                  80                  85 

atc aaa gtc aac ggc acc atg gtg acc aat agc tca cac ctg gaa gtg     1060 
Ile Lys Val Asn Gly Thr Met Val Thr Asn Ser Ser His Leu Glu Val 
 90                  95                 100                 105 

gta aag ctg atc aaa tct ggc gcc tat gtc gca ctc acc ctc ctg ggc     1108 
Val Lys Leu Ile Lys Ser Gly Ala Tyr Val Ala Leu Thr Leu Leu Gly 
                110                 115                 120 

tct tca cct tca tcc atg ggc atc tct ggg ctc cag cag gac cca tcc     1156 
Ser Ser Pro Ser Ser Met Gly Ile Ser Gly Leu Gln Gln Asp Pro Ser 
            125                 130                 135 

cca gca gga gct ccc cga atc acg tca gtg atc ccc tca cca cca cct     1204 
Pro Ala Gly Ala Pro Arg Ile Thr Ser Val Ile Pro Ser Pro Pro Pro 
        140                 145                 150 

cct cca cct cta cca cct cca caa cgc atc aca gga ccc aaa cct ctg     1252 
Pro Pro Pro Leu Pro Pro Pro Gln Arg Ile Thr Gly Pro Lys Pro Leu 
    155                 160                 165 

cag gat ccc gaa gtt caa aaa cat gcc acc cag atc ctc agg aat atg     1300 
Gln Asp Pro Glu Val Gln Lys His Ala Thr Gln Ile Leu Arg Asn Met 
170                 175                 180                 185 

ctg agg cag gaa gaa aaa gaa tta cag gac ata ctt cca cta tat ggt     1348 
Leu Arg Gln Glu Glu Lys Glu Leu Gln Asp Ile Leu Pro Leu Tyr Gly 
                190                 195                 200 

gac acc agc cag aga cca tca gaa ggc cgg ctc tct ctg gat tcc cag     1396 
Asp Thr Ser Gln Arg Pro Ser Glu Gly Arg Leu Ser Leu Asp Ser Gln 
            205                 210                 215 

gag ggg gac agt ggc ttg gac tct ggg aca gaa cgc ttt cct tcc ctc     1444 
Glu Gly Asp Ser Gly Leu Asp Ser Gly Thr Glu Arg Phe Pro Ser Leu 
        220                 225                 230 

agt gag tca ttg atg aat cgg aac tcg gta ctg tca gac cct ggg cta     1492 
Ser Glu Ser Leu Met Asn Arg Asn Ser Val Leu Ser Asp Pro Gly Leu 
    235                 240                 245 

gac agt cct cga acc tcc cct gtg atc atg gcc agg gtg gcc cag cac     1540 
Asp Ser Pro Arg Thr Ser Pro Val Ile Met Ala Arg Val Ala Gln His 
250                 255                 260                 265 

cac agg cgg cag ggc tcg gat gca gca gtc ccc tca acc ggt gac cag     1588 
His Arg Arg Gln Gly Ser Asp Ala Ala Val Pro Ser Thr Gly Asp Gln 
                270                 275                 280 

ggt gta gat caa agc cca aag cct tta att att ggc cca gag gaa gac     1636 
Gly Val Asp Gln Ser Pro Lys Pro Leu Ile Ile Gly Pro Glu Glu Asp 
            285                 290                 295 

tat gac ccg ggt tat ttc aac aac gag agc gac atc ata ttc cag gat     1684 
Tyr Asp Pro Gly Tyr Phe Asn Asn Glu Ser Asp Ile Ile Phe Gln Asp 
        300                 305                 310 

ctg gag aaa ctg aag tct cgg cca gct cac ctg ggg gtt ttt cta cgt     1732 
Leu Glu Lys Leu Lys Ser Arg Pro Ala His Leu Gly Val Phe Leu Arg 
    315                 320                 325 

tac atc ttc tct cag gcg gac ccc agt cca ctg ctt ttt tac ctg tgt     1780 
Tyr Ile Phe Ser Gln Ala Asp Pro Ser Pro Leu Leu Phe Tyr Leu Cys 
330                 335                 340                 345 

gca gaa gtt tat cag cag gca agc ccc aag gat tcc cga agc ttg ggg     1828 
Ala Glu Val Tyr Gln Gln Ala Ser Pro Lys Asp Ser Arg Ser Leu Gly 
                350                 355                 360 

aaa gac atc tgg aat att ttc ctg gag aaa aat gcg cct ctg aga gtg     1876 
Lys Asp Ile Trp Asn Ile Phe Leu Glu Lys Asn Ala Pro Leu Arg Val 
            365                 370                 375 

aag atc cct gag atg cta cag gct gaa att gac tcg cgc ctg cgg aac     1924 
Lys Ile Pro Glu Met Leu Gln Ala Glu Ile Asp Ser Arg Leu Arg Asn 
        380                 385                 390 

agc gaa gat gcc cgt ggt gtt ctc tgt gaa gct caa gag gca gcc atg     1972 
Ser Glu Asp Ala Arg Gly Val Leu Cys Glu Ala Gln Glu Ala Ala Met 
    395                 400                 405 

cct gag atc caa gag cag atc cac gac tac aga acg aag cgc aca ctg     2020 
Pro Glu Ile Gln Glu Gln Ile His Asp Tyr Arg Thr Lys Arg Thr Leu 
410                 415                 420                 425 

ggg ctg ggc agc ctg tat ggt gaa aat gac ctg ctg gac ctg gat ggg     2068 
Gly Leu Gly Ser Leu Tyr Gly Glu Asn Asp Leu Leu Asp Leu Asp Gly 
                430                 435                 440 

gac cct ctc cga gag cgc caa gtg gct gag aag cag ctg gct gcc ctt     2116 
Asp Pro Leu Arg Glu Arg Gln Val Ala Glu Lys Gln Leu Ala Ala Leu 
            445                 450                 455 

gga gat att ttg tcc aag tat gag gaa gac agg agc gcc ccc atg gac     2164 
Gly Asp Ile Leu Ser Lys Tyr Glu Glu Asp Arg Ser Ala Pro Met Asp 
        460                 465                 470 

ttc gcc ctc aat acc tac atg agc cat gct ggg atc cgt ctt cga gag     2212 
Phe Ala Leu Asn Thr Tyr Met Ser His Ala Gly Ile Arg Leu Arg Glu 
    475                 480                 485 

gca cga cct tcc aac aca gct gaa aag gcc cag tct gct cct gac aag     2260 
Ala Arg Pro Ser Asn Thr Ala Glu Lys Ala Gln Ser Ala Pro Asp Lys 
490                 495                 500                 505 

gac aag tgg cta ccg ttc ttc cct aag acc aag aag agc agc aat tcc     2308 
Asp Lys Trp Leu Pro Phe Phe Pro Lys Thr Lys Lys Ser Ser Asn Ser 
                510                 515                 520 

aag aaa gaa aag gat gcc ttg gag gac aag aag cga aac cct atc ctc     2356 
Lys Lys Glu Lys Asp Ala Leu Glu Asp Lys Lys Arg Asn Pro Ile Leu 
            525                 530                 535 

aaa tac att ggg aag ccc aaa agc tct tct caa agc aca ttt cat att     2404 
Lys Tyr Ile Gly Lys Pro Lys Ser Ser Ser Gln Ser Thr Phe His Ile 
        540                 545                 550 

ccc ttg tcc cct gtg gaa gtc aaa cca ggc aat gtg agg aac atc att     2452 
Pro Leu Ser Pro Val Glu Val Lys Pro Gly Asn Val Arg Asn Ile Ile 
    555                 560                 565 

cag cac ttt gag aac aac cag cag tat gat gcc cca gaa cct ggg aca     2500 
Gln His Phe Glu Asn Asn Gln Gln Tyr Asp Ala Pro Glu Pro Gly Thr 
570                 575                 580                 585 

caa cga ctc tcg acc gga agc ttt cct gag gac ctg ctg gag agt gac     2548 
Gln Arg Leu Ser Thr Gly Ser Phe Pro Glu Asp Leu Leu Glu Ser Asp 
                590                 595                 600 

agt tca cgc tca gag att cgc ctg ggc cgc tct gaa agc ctc aag ggc     2596 
Ser Ser Arg Ser Glu Ile Arg Leu Gly Arg Ser Glu Ser Leu Lys Gly 
            605                 610                 615 

cgg gaa gag atg aaa cgg tct cga aag gca gag aac gtg ccc cgc tct     2644 
Arg Glu Glu Met Lys Arg Ser Arg Lys Ala Glu Asn Val Pro Arg Ser 
        620                 625                 630 

cgc agt gat gtt gac atg gat gct gct gcg gag gct act cgc ctg cac     2692 
Arg Ser Asp Val Asp Met Asp Ala Ala Ala Glu Ala Thr Arg Leu His 
    635                 640                 645 

cag tca gcc tcg tcc tct acc tcc agc ctc tcc acc agg tct ctt gag     2740 
Gln Ser Ala Ser Ser Ser Thr Ser Ser Leu Ser Thr Arg Ser Leu Glu 
650                 655                 660                 665 

aac cca acc cct cca ttc act ccc aaa atg ggc cgc agg agc att gag     2788 
Asn Pro Thr Pro Pro Phe Thr Pro Lys Met Gly Arg Arg Ser Ile Glu 
                670                 675                 680 

tcc ccc agt ttg ggg ttc tgc aca gat acc ctc ctt ccc cac ctc cta     2836 
Ser Pro Ser Leu Gly Phe Cys Thr Asp Thr Leu Leu Pro His Leu Leu 
            685                 690                 695 

gag gat gat ctg ggc cag ctg tct gac ctg gag cca gag cca gat gcc     2884 
Glu Asp Asp Leu Gly Gln Leu Ser Asp Leu Glu Pro Glu Pro Asp Ala 
        700                 705                 710 

caa aat tgg cag cat aca gtg ggc aag gat gtg gtg gct ggg cta acc     2932 
Gln Asn Trp Gln His Thr Val Gly Lys Asp Val Val Ala Gly Leu Thr 
    715                 720                 725 

cag cgg gag att gac cgg caa gag gtc atc aat gag ctg ttt gtg act     2980 
Gln Arg Glu Ile Asp Arg Gln Glu Val Ile Asn Glu Leu Phe Val Thr 
730                 735                 740                 745 

gaa gct tcc cac ctg cgc aca ctc cgg gtc ctg gac ctg atc ttc tac     3028 
Glu Ala Ser His Leu Arg Thr Leu Arg Val Leu Asp Leu Ile Phe Tyr 
                750                 755                 760 

cag cga atg aag aag gag aac ctg atg ccc cgg gag gag ctg gcc cgg     3076 
Gln Arg Met Lys Lys Glu Asn Leu Met Pro Arg Glu Glu Leu Ala Arg 
            765                 770                 775 

ctc ttc ccg aac ctg cct gaa ctc ata gag att cac aat tcc tgg tgt     3124 
Leu Phe Pro Asn Leu Pro Glu Leu Ile Glu Ile His Asn Ser Trp Cys 
        780                 785                 790 

gaa gcc atg aag aag ctc cgg gag gaa ggc ccc atc atc aaa gag atc     3172 
Glu Ala Met Lys Lys Leu Arg Glu Glu Gly Pro Ile Ile Lys Glu Ile 
    795                 800                 805 

agt gac ctc atg ctg gcc cgg ttt gat ggc cct gcc cga gag gaa ctc     3220 
Ser Asp Leu Met Leu Ala Arg Phe Asp Gly Pro Ala Arg Glu Glu Leu 
810                 815                 820                 825 

cag caa gtg gct gca cag ttc tgt tcc tat cag tca ata gcc cta gag     3268 
Gln Gln Val Ala Ala Gln Phe Cys Ser Tyr Gln Ser Ile Ala Leu Glu 
                830                 835                 840 

cta atc aag acc aag caa cgc aag gag agt cga ttc cag ctc ttc atg     3316 
Leu Ile Lys Thr Lys Gln Arg Lys Glu Ser Arg Phe Gln Leu Phe Met 
            845                 850                 855 

cag gag gct gag agc cac cct cag tgt cgg cgg ctg cag ctg aga gac     3364 
Gln Glu Ala Glu Ser His Pro Gln Cys Arg Arg Leu Gln Leu Arg Asp 
        860                 865                 870 

ctc atc atc tct gag atg cag cgg ctc acc aag tac ccg ctg ctg ctg     3412 
Leu Ile Ile Ser Glu Met Gln Arg Leu Thr Lys Tyr Pro Leu Leu Leu 
    875                 880                 885 

gag agc atc atc aag cac aca gag ggt ggc acc tct gag cat gag aag     3460 
Glu Ser Ile Ile Lys His Thr Glu Gly Gly Thr Ser Glu His Glu Lys 
890                 895                 900                 905 

ctg tgc cgg gcc cgg gac cag tgc cgg gag att ctc aag tat gtg aat     3508 
Leu Cys Arg Ala Arg Asp Gln Cys Arg Glu Ile Leu Lys Tyr Val Asn 
                910                 915                 920 

gaa gcg gta aaa caa aca gag aac cgc cac cgt tta gag ggc tac cag     3556 
Glu Ala Val Lys Gln Thr Glu Asn Arg His Arg Leu Glu Gly Tyr Gln 
            925                 930                 935 

aaa cgc ctg gat gcc acc gcc ctg gag agg gcc agc aac ccc ctg gca     3604 
Lys Arg Leu Asp Ala Thr Ala Leu Glu Arg Ala Ser Asn Pro Leu Ala 
        940                 945                 950 

gca gag ttc aag agc ctg gat ctt aca acc aga aaa atg atc cat gag     3652 
Ala Glu Phe Lys Ser Leu Asp Leu Thr Thr Arg Lys Met Ile His Glu 
    955                 960                 965 

gga ccc ctg acc tgg agg atc agc aag gat aag acc ttg gac ctc cac     3700 
Gly Pro Leu Thr Trp Arg Ile Ser Lys Asp Lys Thr Leu Asp Leu His 
970                 975                 980                 985 

gtg ctg ctg ctg gag gac ctc cta gtg ctg cta cag aaa cag gat gag     3748 
Val Leu Leu Leu Glu Asp Leu Leu Val Leu Leu Gln Lys Gln Asp Glu 
                990                 995                1000 

aag cta ttg ctg aag tgc cac agc aag act gct gtg ggc tcc tca gac     3796 
Lys Leu Leu Leu Lys Cys His Ser Lys Thr Ala Val Gly Ser Ser Asp 
           1005                1010                1015 

agc aag cag acc ttc agc ccc gtg ctc aag ctc aat gct gtg ctc atc     3844 
Ser Lys Gln Thr Phe Ser Pro Val Leu Lys Leu Asn Ala Val Leu Ile 
       1020                1025                1030 

cgc tct gtg gcc aca gat aaa cgg gcc ttc ttc atc atc tgc acc tcc     3892 
Arg Ser Val Ala Thr Asp Lys Arg Ala Phe Phe Ile Ile Cys Thr Ser 
   1035                1040                1045 

aag ctg ggc cca ccc cag atc tat gag ctg gtt gca ttg acg tca tca     3940 
Lys Leu Gly Pro Pro Gln Ile Tyr Glu Leu Val Ala Leu Thr Ser Ser 
1050               1055                1060                1065 

gac aag aac aca tgg atg gag ctc tta gaa gag gcc gtg cgg aat gcc     3988 
Asp Lys Asn Thr Trp Met Glu Leu Leu Glu Glu Ala Val Arg Asn Ala 
               1070                1075                1080 

acc agg cac ccc gga gct gcc cca atg ccc gtc cat cct cca ccc cca     4036 
Thr Arg His Pro Gly Ala Ala Pro Met Pro Val His Pro Pro Pro Pro 
           1085                1090                1095 

ggt ccc cgg gag cca gcc cag cag ggc ccc aca ccc agc agg gta gaa     4084 
Gly Pro Arg Glu Pro Ala Gln Gln Gly Pro Thr Pro Ser Arg Val Glu 
       1100                1105                1110 

ctg gat gac tca gac gtg ttc cat ggt gaa cct gaa cct gag gag ctg     4132 
Leu Asp Asp Ser Asp Val Phe His Gly Glu Pro Glu Pro Glu Glu Leu 
   1115                1120                1125 

cct gga ggc act ggg tcc cag cag agg gtc caa ggg aag cac cag gtc     4180 
Pro Gly Gly Thr Gly Ser Gln Gln Arg Val Gln Gly Lys His Gln Val 
1130               1135                1140                1145 

ctg cta gag gac cct gag cag gag ggc agt gca gag gaa gag gaa ctg     4228 
Leu Leu Glu Asp Pro Glu Gln Glu Gly Ser Ala Glu Glu Glu Glu Leu 
               1150                1155                1160 

ggt gtc ctg cct tgc cct tcc aca tcc ctg gat gga gag aac agg ggc     4276 
Gly Val Leu Pro Cys Pro Ser Thr Ser Leu Asp Gly Glu Asn Arg Gly 
           1165                1170                1175 

atc agg aca agg aac ccc atc cac ttg gcc ttc cca ggc cct ctg ttc     4324 
Ile Arg Thr Arg Asn Pro Ile His Leu Ala Phe Pro Gly Pro Leu Phe 
       1180                1185                1190 

atg gaa ggg ctc gct gac tcc gct ctg gaa gat gtg gag aac ctg cga     4372 
Met Glu Gly Leu Ala Asp Ser Ala Leu Glu Asp Val Glu Asn Leu Arg 
   1195                1200                1205 

cat ctg atc ctg tgg agc ctg ctg cca ggt cac acc atg gaa act cag     4420 
His Leu Ile Leu Trp Ser Leu Leu Pro Gly His Thr Met Glu Thr Gln 
1210               1215                1220                1225 

gct gcc cag gag ccc gag gac gac ctg aca ccc aca cct tct gtc atc     4468 
Ala Ala Gln Glu Pro Glu Asp Asp Leu Thr Pro Thr Pro Ser Val Ile 
               1230                1235                1240 

agc gtc acc tct cac ccc tgg gac cca ggc tcc cca ggg caa gca ccc     4516 
Ser Val Thr Ser His Pro Trp Asp Pro Gly Ser Pro Gly Gln Ala Pro 
           1245                1250                1255 

cct ggg ggt gaa ggg gac aac acc cag ctt gca ggg ctg gag ggg gaa     4564 
Pro Gly Gly Glu Gly Asp Asn Thr Gln Leu Ala Gly Leu Glu Gly Glu 
       1260                1265                1270 

cgg cca gag cag gaa gac atg ggt ctc tgt tct ctg gaa cac cta ccc     4612 
Arg Pro Glu Gln Glu Asp Met Gly Leu Cys Ser Leu Glu His Leu Pro 
   1275                1280                1285 

cca agg acc aga aat tct ggg ata tgg gag tct cca gaa ctg gac agg     4660 
Pro Arg Thr Arg Asn Ser Gly Ile Trp Glu Ser Pro Glu Leu Asp Arg 
1290               1295                1300                1305 

aat ctg gct gaa gat gct tca agc aca gag gca gca gga ggt tac aaa     4708 
Asn Leu Ala Glu Asp Ala Ser Ser Thr Glu Ala Ala Gly Gly Tyr Lys 
               1310                1315                1320 

gtt gtg aga aaa gct gag gtg gca ggc agc aag gtt gtc cct gca cta     4756 
Val Val Arg Lys Ala Glu Val Ala Gly Ser Lys Val Val Pro Ala Leu 
           1325                1330                1335 

cca gag agt ggc cag tca gag cct ggg cca cct gaa gtg gaa ggc gga     4804 
Pro Glu Ser Gly Gln Ser Glu Pro Gly Pro Pro Glu Val Glu Gly Gly 
       1340                1345                1350 

aca aag gct acg ggg aac tgc ttt tat gtc agc atg cca tca gga ccc     4852 
Thr Lys Ala Thr Gly Asn Cys Phe Tyr Val Ser Met Pro Ser Gly Pro 
   1355                1360                1365 

ccg gac tca agc acc gac cac tca gag gca ccc atg agc ccc cct cag     4900 
Pro Asp Ser Ser Thr Asp His Ser Glu Ala Pro Met Ser Pro Pro Gln 
1370               1375                1380                1385 

cct gac agc ctc cct gca ggg cag aca gag cct cag cct cag ctg cag     4948 
Pro Asp Ser Leu Pro Ala Gly Gln Thr Glu Pro Gln Pro Gln Leu Gln 
               1390                1395                1400 

gga ggc aac gat gat cca aga cgc ccc agc cgc tct cct cca agc ctg     4996 
Gly Gly Asn Asp Asp Pro Arg Arg Pro Ser Arg Ser Pro Pro Ser Leu 
           1405                1410                1415 

gcc ctc agg gac gtg ggc atg atc ttc cat acc att gag cag ctc act     5044 
Ala Leu Arg Asp Val Gly Met Ile Phe His Thr Ile Glu Gln Leu Thr 
       1420                1425                1430 

ctc aag ctc aac agg ctc aag gat atg gag ctg gcc cac aga gag ctg     5092 
Leu Lys Leu Asn Arg Leu Lys Asp Met Glu Leu Ala His Arg Glu Leu 
   1435                1440                1445 

ctc aag tcc ctt ggg gga gag tca tct ggt ggc acc acg cct gtg ggc     5140 
Leu Lys Ser Leu Gly Gly Glu Ser Ser Gly Gly Thr Thr Pro Val Gly 
1450               1455                1460                1465 

agt ttc cac aca gaa gca gct aga tgg aca gat ggc tcc ctc tca cct     5188 
Ser Phe His Thr Glu Ala Ala Arg Trp Thr Asp Gly Ser Leu Ser Pro 
               1470                1475                1480 

ccc gct aag gag ccc cta gct tct gac tcc agg aac agc cat gaa ctg     5236 
Pro Ala Lys Glu Pro Leu Ala Ser Asp Ser Arg Asn Ser His Glu Leu 
           1485                1490                1495 

ggg ccc tgc cct gag gat ggc tct gac gcc ccc ctg gaa gac agc aca     5284 
Gly Pro Cys Pro Glu Asp Gly Ser Asp Ala Pro Leu Glu Asp Ser Thr 
       1500                1505                1510 

gca gac gca gcc gcg tca cca gga cca taa ccgtacaaac caccaaatcc       5334 
Ala Asp Ala Ala Ala Ser Pro Gly Pro 
   1515                1520 

tctgcgtccc cactcctcct tcagggactg gcctgagacc ggggcacagg gtagggggga   5394 

tcccaacact cctccctgtg gaggaggcag ttagggaaac taggatccag ccaaggcccg   5454 

gggggagacc cgcatgttgc ttggtctgct caagtcggag tcaggtttca gtgtcttttc   5514 

cctcccttag cccaaccctc caaggcctca tgtctcctaa gcatgctgac tgcatccgaa   5574 

aggcccccac tcaccatggt ctgccctcac cccacatatg tgtgtacacg cgcacgcctg   5634 

tatgtgcgct gccctcagac atgcaagtga aaggaggagg cttctgtgta aatgcacttt   5694 

cttcctcccc tctttctcca taagacccca ggcagaggtg ggtgcctccc ctcccctctt   5754 

tgtcactttg gtttcctata aatatgtatg tatcgt                             5790 

 
           
             8  
             1522  
             PRT  
             Homo sapiens  
           
            8 

Met Ser Val Arg Leu Pro Gln Ser Ile Asp Arg Leu Ser Ser Leu Ser 
  1               5                  10                  15 

Ser Leu Gly Asp Ser Ala Pro Glu Arg Lys Ser Pro Ser His His Arg 
             20                  25                  30 

Gln Pro Ser Asp Ala Ser Glu Thr Thr Gly Leu Val Gln Arg Cys Val 
         35                  40                  45 

Ile Ile Gln Lys Asp Gln His Gly Phe Gly Phe Thr Val Ser Gly Asp 
     50                  55                  60 

Arg Ile Val Leu Val Gln Ser Val Arg Pro Gly Gly Ala Ala Met Lys 
 65                  70                  75                  80 

Ala Gly Val Lys Glu Gly Asp Arg Ile Ile Lys Val Asn Gly Thr Met 
                 85                  90                  95 

Val Thr Asn Ser Ser His Leu Glu Val Val Lys Leu Ile Lys Ser Gly 
            100                 105                 110 

Ala Tyr Val Ala Leu Thr Leu Leu Gly Ser Ser Pro Ser Ser Met Gly 
        115                 120                 125 

Ile Ser Gly Leu Gln Gln Asp Pro Ser Pro Ala Gly Ala Pro Arg Ile 
    130                 135                 140 

Thr Ser Val Ile Pro Ser Pro Pro Pro Pro Pro Pro Leu Pro Pro Pro 
145                 150                 155                 160 

Gln Arg Ile Thr Gly Pro Lys Pro Leu Gln Asp Pro Glu Val Gln Lys 
                165                 170                 175 

His Ala Thr Gln Ile Leu Arg Asn Met Leu Arg Gln Glu Glu Lys Glu 
            180                 185                 190 

Leu Gln Asp Ile Leu Pro Leu Tyr Gly Asp Thr Ser Gln Arg Pro Ser 
        195                 200                 205 

Glu Gly Arg Leu Ser Leu Asp Ser Gln Glu Gly Asp Ser Gly Leu Asp 
    210                 215                 220 

Ser Gly Thr Glu Arg Phe Pro Ser Leu Ser Glu Ser Leu Met Asn Arg 
225                 230                 235                 240 

Asn Ser Val Leu Ser Asp Pro Gly Leu Asp Ser Pro Arg Thr Ser Pro 
                245                 250                 255 

Val Ile Met Ala Arg Val Ala Gln His His Arg Arg Gln Gly Ser Asp 
            260                 265                 270 

Ala Ala Val Pro Ser Thr Gly Asp Gln Gly Val Asp Gln Ser Pro Lys 
        275                 280                 285 

Pro Leu Ile Ile Gly Pro Glu Glu Asp Tyr Asp Pro Gly Tyr Phe Asn 
    290                 295                 300 

Asn Glu Ser Asp Ile Ile Phe Gln Asp Leu Glu Lys Leu Lys Ser Arg 
305                 310                 315                 320 

Pro Ala His Leu Gly Val Phe Leu Arg Tyr Ile Phe Ser Gln Ala Asp 
                325                 330                 335 

Pro Ser Pro Leu Leu Phe Tyr Leu Cys Ala Glu Val Tyr Gln Gln Ala 
            340                 345                 350 

Ser Pro Lys Asp Ser Arg Ser Leu Gly Lys Asp Ile Trp Asn Ile Phe 
        355                 360                 365 

Leu Glu Lys Asn Ala Pro Leu Arg Val Lys Ile Pro Glu Met Leu Gln 
    370                 375                 380 

Ala Glu Ile Asp Ser Arg Leu Arg Asn Ser Glu Asp Ala Arg Gly Val 
385                 390                 395                 400 

Leu Cys Glu Ala Gln Glu Ala Ala Met Pro Glu Ile Gln Glu Gln Ile 
                405                 410                 415 

His Asp Tyr Arg Thr Lys Arg Thr Leu Gly Leu Gly Ser Leu Tyr Gly 
            420                 425                 430 

Glu Asn Asp Leu Leu Asp Leu Asp Gly Asp Pro Leu Arg Glu Arg Gln 
        435                 440                 445 

Val Ala Glu Lys Gln Leu Ala Ala Leu Gly Asp Ile Leu Ser Lys Tyr 
    450                 455                 460 

Glu Glu Asp Arg Ser Ala Pro Met Asp Phe Ala Leu Asn Thr Tyr Met 
465                 470                 475                 480 

Ser His Ala Gly Ile Arg Leu Arg Glu Ala Arg Pro Ser Asn Thr Ala 
                485                 490                 495 

Glu Lys Ala Gln Ser Ala Pro Asp Lys Asp Lys Trp Leu Pro Phe Phe 
            500                 505                 510 

Pro Lys Thr Lys Lys Ser Ser Asn Ser Lys Lys Glu Lys Asp Ala Leu 
        515                 520                 525 

Glu Asp Lys Lys Arg Asn Pro Ile Leu Lys Tyr Ile Gly Lys Pro Lys 
    530                 535                 540 

Ser Ser Ser Gln Ser Thr Phe His Ile Pro Leu Ser Pro Val Glu Val 
545                 550                 555                 560 

Lys Pro Gly Asn Val Arg Asn Ile Ile Gln His Phe Glu Asn Asn Gln 
                565                 570                 575 

Gln Tyr Asp Ala Pro Glu Pro Gly Thr Gln Arg Leu Ser Thr Gly Ser 
            580                 585                 590 

Phe Pro Glu Asp Leu Leu Glu Ser Asp Ser Ser Arg Ser Glu Ile Arg 
        595                 600                 605 

Leu Gly Arg Ser Glu Ser Leu Lys Gly Arg Glu Glu Met Lys Arg Ser 
    610                 615                 620 

Arg Lys Ala Glu Asn Val Pro Arg Ser Arg Ser Asp Val Asp Met Asp 
625                 630                 635                 640 

Ala Ala Ala Glu Ala Thr Arg Leu His Gln Ser Ala Ser Ser Ser Thr 
                645                 650                 655 

Ser Ser Leu Ser Thr Arg Ser Leu Glu Asn Pro Thr Pro Pro Phe Thr 
            660                 665                 670 

Pro Lys Met Gly Arg Arg Ser Ile Glu Ser Pro Ser Leu Gly Phe Cys 
        675                 680                 685 

Thr Asp Thr Leu Leu Pro His Leu Leu Glu Asp Asp Leu Gly Gln Leu 
    690                 695                 700 

Ser Asp Leu Glu Pro Glu Pro Asp Ala Gln Asn Trp Gln His Thr Val 
705                 710                 715                 720 

Gly Lys Asp Val Val Ala Gly Leu Thr Gln Arg Glu Ile Asp Arg Gln 
                725                 730                 735 

Glu Val Ile Asn Glu Leu Phe Val Thr Glu Ala Ser His Leu Arg Thr 
            740                 745                 750 

Leu Arg Val Leu Asp Leu Ile Phe Tyr Gln Arg Met Lys Lys Glu Asn 
        755                 760                 765 

Leu Met Pro Arg Glu Glu Leu Ala Arg Leu Phe Pro Asn Leu Pro Glu 
    770                 775                 780 

Leu Ile Glu Ile His Asn Ser Trp Cys Glu Ala Met Lys Lys Leu Arg 
785                 790                 795                 800 

Glu Glu Gly Pro Ile Ile Lys Glu Ile Ser Asp Leu Met Leu Ala Arg 
                805                 810                 815 

Phe Asp Gly Pro Ala Arg Glu Glu Leu Gln Gln Val Ala Ala Gln Phe 
            820                 825                 830 

Cys Ser Tyr Gln Ser Ile Ala Leu Glu Leu Ile Lys Thr Lys Gln Arg 
        835                 840                 845 

Lys Glu Ser Arg Phe Gln Leu Phe Met Gln Glu Ala Glu Ser His Pro 
    850                 855                 860 

Gln Cys Arg Arg Leu Gln Leu Arg Asp Leu Ile Ile Ser Glu Met Gln 
865                 870                 875                 880 

Arg Leu Thr Lys Tyr Pro Leu Leu Leu Glu Ser Ile Ile Lys His Thr 
                885                 890                 895 

Glu Gly Gly Thr Ser Glu His Glu Lys Leu Cys Arg Ala Arg Asp Gln 
            900                 905                 910 

Cys Arg Glu Ile Leu Lys Tyr Val Asn Glu Ala Val Lys Gln Thr Glu 
        915                 920                 925 

Asn Arg His Arg Leu Glu Gly Tyr Gln Lys Arg Leu Asp Ala Thr Ala 
    930                 935                 940 

Leu Glu Arg Ala Ser Asn Pro Leu Ala Ala Glu Phe Lys Ser Leu Asp 
945                 950                 955                 960 

Leu Thr Thr Arg Lys Met Ile His Glu Gly Pro Leu Thr Trp Arg Ile 
                965                 970                 975 

Ser Lys Asp Lys Thr Leu Asp Leu His Val Leu Leu Leu Glu Asp Leu 
            980                 985                 990 

Leu Val Leu Leu Gln Lys Gln Asp Glu Lys Leu Leu Leu Lys Cys His 
        995                1000                1005 

Ser Lys Thr Ala Val Gly Ser Ser Asp Ser Lys Gln Thr Phe Ser Pro 
   1010                1015                1020 

Val Leu Lys Leu Asn Ala Val Leu Ile Arg Ser Val Ala Thr Asp Lys 
1025                1030                1035               1040 

Arg Ala Phe Phe Ile Ile Cys Thr Ser Lys Leu Gly Pro Pro Gln Ile 
               1045                1050                1055 

Tyr Glu Leu Val Ala Leu Thr Ser Ser Asp Lys Asn Thr Trp Met Glu 
           1060                1065                1070 

Leu Leu Glu Glu Ala Val Arg Asn Ala Thr Arg His Pro Gly Ala Ala 
       1075                1080                1085 

Pro Met Pro Val His Pro Pro Pro Pro Gly Pro Arg Glu Pro Ala Gln 
   1090                1095                1100 

Gln Gly Pro Thr Pro Ser Arg Val Glu Leu Asp Asp Ser Asp Val Phe 
1105               1110                1115                1120 

His Gly Glu Pro Glu Pro Glu Glu Leu Pro Gly Gly Thr Gly Ser Gln 
               1125                1130                1135 

Gln Arg Val Gln Gly Lys His Gln Val Leu Leu Glu Asp Pro Glu Gln 
           1140                1145                1150 

Glu Gly Ser Ala Glu Glu Glu Glu Leu Gly Val Leu Pro Cys Pro Ser 
       1155                1160                1165 

Thr Ser Leu Asp Gly Glu Asn Arg Gly Ile Arg Thr Arg Asn Pro Ile 
   1170                1175                1180 

His Leu Ala Phe Pro Gly Pro Leu Phe Met Glu Gly Leu Ala Asp Ser 
1185               1190                1195                1200 

Ala Leu Glu Asp Val Glu Asn Leu Arg His Leu Ile Leu Trp Ser Leu 
               1205                1210                1215 

Leu Pro Gly His Thr Met Glu Thr Gln Ala Ala Gln Glu Pro Glu Asp 
           1220                1225                1230 

Asp Leu Thr Pro Thr Pro Ser Val Ile Ser Val Thr Ser His Pro Trp 
       1235                1240                1245 

Asp Pro Gly Ser Pro Gly Gln Ala Pro Pro Gly Gly Glu Gly Asp Asn 
   1250                1255                1260 

Thr Gln Leu Ala Gly Leu Glu Gly Glu Arg Pro Glu Gln Glu Asp Met 
1265               1270                1275                1280 

Gly Leu Cys Ser Leu Glu His Leu Pro Pro Arg Thr Arg Asn Ser Gly 
               1285                1290                1295 

Ile Trp Glu Ser Pro Glu Leu Asp Arg Asn Leu Ala Glu Asp Ala Ser 
           1300                1305                1310 

Ser Thr Glu Ala Ala Gly Gly Tyr Lys Val Val Arg Lys Ala Glu Val 
       1315                1320                1325 

Ala Gly Ser Lys Val Val Pro Ala Leu Pro Glu Ser Gly Gln Ser Glu 
   1330                1335                1340 

Pro Gly Pro Pro Glu Val Glu Gly Gly Thr Lys Ala Thr Gly Asn Cys 
1345               1350                1355                1360 

Phe Tyr Val Ser Met Pro Ser Gly Pro Pro Asp Ser Ser Thr Asp His 
               1365                1370                1375 

Ser Glu Ala Pro Met Ser Pro Pro Gln Pro Asp Ser Leu Pro Ala Gly 
           1380                1385                1390 

Gln Thr Glu Pro Gln Pro Gln Leu Gln Gly Gly Asn Asp Asp Pro Arg 
       1395                1400                1405 

Arg Pro Ser Arg Ser Pro Pro Ser Leu Ala Leu Arg Asp Val Gly Met 
   1410                1415                1420 

Ile Phe His Thr Ile Glu Gln Leu Thr Leu Lys Leu Asn Arg Leu Lys 
1425               1430                1435                1440 

Asp Met Glu Leu Ala His Arg Glu Leu Leu Lys Ser Leu Gly Gly Glu 
               1445                1450                1455 

Ser Ser Gly Gly Thr Thr Pro Val Gly Ser Phe His Thr Glu Ala Ala 
           1460                1465                1470 

Arg Trp Thr Asp Gly Ser Leu Ser Pro Pro Ala Lys Glu Pro Leu Ala 
       1475                1480                1485 

Ser Asp Ser Arg Asn Ser His Glu Leu Gly Pro Cys Pro Glu Asp Gly 
   1490                1495                1500 

Ser Asp Ala Pro Leu Glu Asp Ser Thr Ala Asp Ala Ala Ala Ser Pro 
1505               1510                1515                1520 

Gly Pro 

 
           
             9  
             2215  
             DNA  
             Homo sapiens  
             
               CDS  
               (242)..(1783)  
             
           
            9 

actcagtaga ccgccactgg ctgtgcacgt tatggggttt ccacctaggg ctcggcctga     60 

ggcttgtaac actccgtttt cccccgagtc acaggggcag tcttgcccct cgcagctggg    120 

tcgcggtgtc tctcaaaggt ccccctctac aggggcttcg tgaggcccgg gcccacaggg    180 

cgctcggtcc cggaagtgac gtctcccaga ggggccggaa gtggcagtgg agggagggaa    240 

g atg gcg gag gtg ggg gag ata atc gag ggc tgc cgc cta ccc gtg ctg    289 
  Met Ala Glu Val Gly Glu Ile Ile Glu Gly Cys Arg Leu Pro Val Leu 
    1               5                  10                  15 

cgg cgg aac cag gac aac gaa gat gag tgg ccc ctg gcc gag atc ctg      337 
Arg Arg Asn Gln Asp Asn Glu Asp Glu Trp Pro Leu Ala Glu Ile Leu 
             20                  25                  30 

agc gtg aag gac atc agt ggc cgg aag ctt ttc tac gtc cat tac att      385 
Ser Val Lys Asp Ile Ser Gly Arg Lys Leu Phe Tyr Val His Tyr Ile 
         35                  40                  45 

gac ttc aac aaa cgt ctg gat gaa tgg gtg acg cat gag cgg ctg gac      433 
Asp Phe Asn Lys Arg Leu Asp Glu Trp Val Thr His Glu Arg Leu Asp 
     50                  55                  60 

cta aag aag atc cag ttc ccc aag aaa gag gcc aag acc ccc act aag      481 
Leu Lys Lys Ile Gln Phe Pro Lys Lys Glu Ala Lys Thr Pro Thr Lys 
 65                  70                  75                  80 

aac gga ctt cct ggg tcc cgt cct ggc tct cca gag aga gag gtg ccg      529 
Asn Gly Leu Pro Gly Ser Arg Pro Gly Ser Pro Glu Arg Glu Val Pro 
                 85                  90                  95 

gcc tcg gcg cag gcc agc ggg aag acc ttg cca atc ccg gtc cag atc      577 
Ala Ser Ala Gln Ala Ser Gly Lys Thr Leu Pro Ile Pro Val Gln Ile 
            100                 105                 110 

aca ctc cgc ttc aac ctg ccc aag gag cgg gag gcc att ccc ggt ggc      625 
Thr Leu Arg Phe Asn Leu Pro Lys Glu Arg Glu Ala Ile Pro Gly Gly 
        115                 120                 125 

gag cct gac cag ccg ctc tcc tcc agc tcc tgc ctg cag ccc aac cac      673 
Glu Pro Asp Gln Pro Leu Ser Ser Ser Ser Cys Leu Gln Pro Asn His 
    130                 135                 140 

cgc tca acg aaa cgg aag gtg gag gtg gtt tca cca gca act cca gtg      721 
Arg Ser Thr Lys Arg Lys Val Glu Val Val Ser Pro Ala Thr Pro Val 
145                 150                 155                 160 

ccc agc gag aca gcc ccg gcc tcg gtt ttt ccc cag aat gga gcc gcc      769 
Pro Ser Glu Thr Ala Pro Ala Ser Val Phe Pro Gln Asn Gly Ala Ala 
                165                 170                 175 

cgt agg gca gtg gca gcc cag cca gga cgg aag cga aaa tcg aat tgt      817 
Arg Arg Ala Val Ala Ala Gln Pro Gly Arg Lys Arg Lys Ser Asn Cys 
            180                 185                 190 

ttg ggc act gat gag gac tcc cag gac agc tct gat gga ata ccg tca      865 
Leu Gly Thr Asp Glu Asp Ser Gln Asp Ser Ser Asp Gly Ile Pro Ser 
        195                 200                 205 

gca cca cgc atg act ggc agc ctg gtg tct gat cga agc cac gac gac      913 
Ala Pro Arg Met Thr Gly Ser Leu Val Ser Asp Arg Ser His Asp Asp 
    210                 215                 220 

atc gtc acc cgg atg aag aac att gag tgc att gag ctg ggc cgg cac      961 
Ile Val Thr Arg Met Lys Asn Ile Glu Cys Ile Glu Leu Gly Arg His 
225                 230                 235                 240 

cgc ctc aag ccg tgg tac ttc tcc ccg tac cca cag gaa ctc acc aca     1009 
Arg Leu Lys Pro Trp Tyr Phe Ser Pro Tyr Pro Gln Glu Leu Thr Thr 
                245                 250                 255 

ttg cct gtc ctc tac ctg tgc gag ttc tgc ctc aag tac ggc cgt agt     1057 
Leu Pro Val Leu Tyr Leu Cys Glu Phe Cys Leu Lys Tyr Gly Arg Ser 
            260                 265                 270 

ctc aag tgt ctt cag cgt cat ttg acc aag tgt gac cta cga cat cct     1105 
Leu Lys Cys Leu Gln Arg His Leu Thr Lys Cys Asp Leu Arg His Pro 
        275                 280                 285 

cca ggc aat gag att tac cgc aag ggc acc atc tcc ttc ttt gag att     1153 
Pro Gly Asn Glu Ile Tyr Arg Lys Gly Thr Ile Ser Phe Phe Glu Ile 
    290                 295                 300 

gat gga cgt aag aac aag agt tat tcc cag aac ctg tgt ctt ttg gcc     1201 
Asp Gly Arg Lys Asn Lys Ser Tyr Ser Gln Asn Leu Cys Leu Leu Ala 
305                 310                 315                 320 

aag tgt ttc ctt gac cat aag aca ctg tac tat gac aca gac cct ttc     1249 
Lys Cys Phe Leu Asp His Lys Thr Leu Tyr Tyr Asp Thr Asp Pro Phe 
                325                 330                 335 

ctc ttc tac gtc atg aca gag tat gac tgt aag ggc ttc cac atc gtg     1297 
Leu Phe Tyr Val Met Thr Glu Tyr Asp Cys Lys Gly Phe His Ile Val 
            340                 345                 350 

ggc tac ttc tcc aag gag aaa gaa tca acg gaa gac tac aat gtg gcc     1345 
Gly Tyr Phe Ser Lys Glu Lys Glu Ser Thr Glu Asp Tyr Asn Val Ala 
        355                 360                 365 

tgc atc cta acc ctg cct ccc tac cag cgc cgg ggc tac cgg aag ctg     1393 
Cys Ile Leu Thr Leu Pro Pro Tyr Gln Arg Arg Gly Tyr Arg Lys Leu 
    370                 375                 380 

ctg atc gag ttc agc tat gaa ctc tcc aaa gtg gaa ggg aaa aca ggg     1441 
Leu Ile Glu Phe Ser Tyr Glu Leu Ser Lys Val Glu Gly Lys Thr Gly 
385                 390                 395                 400 

acc cct gag aag ccc ctc tca gac ctt ggc ctc cta tcc tat cga agc     1489 
Thr Pro Glu Lys Pro Leu Ser Asp Leu Gly Leu Leu Ser Tyr Arg Ser 
                405                 410                 415 

tac tgg tcc cag acc atc ctg gag atc ctg atg ggg ctg aag tcg gag     1537 
Tyr Trp Ser Gln Thr Ile Leu Glu Ile Leu Met Gly Leu Lys Ser Glu 
            420                 425                 430 

agc ggg gag agg cca cag atc acc atc aat gag att agt gaa atc acc     1585 
Ser Gly Glu Arg Pro Gln Ile Thr Ile Asn Glu Ile Ser Glu Ile Thr 
        435                 440                 445 

agc atc aag aag gag gat gtc atc tcc act ctg cag tac ctc aat ctc     1633 
Ser Ile Lys Lys Glu Asp Val Ile Ser Thr Leu Gln Tyr Leu Asn Leu 
    450                 455                 460 

atc aac tac tac aag ggc cag tac atc ctc aca ctg tca gag gac atc     1681 
Ile Asn Tyr Tyr Lys Gly Gln Tyr Ile Leu Thr Leu Ser Glu Asp Ile 
465                 470                 475                 480 

gtg gat ggc cat gag cgg gcc atg ctc aag cgg ctc ctg cgg atc gac     1729 
Val Asp Gly His Glu Arg Ala Met Leu Lys Arg Leu Leu Arg Ile Asp 
                485                 490                 495 

tcc aag tgt ctg cac ttc act ccc aag gac tgg agc aag agg ggg aag     1777 
Ser Lys Cys Leu His Phe Thr Pro Lys Asp Trp Ser Lys Arg Gly Lys 
            500                 505                 510 

tgg tga ccagacactg cccactgcag tgccaagacg gcagcaggac tggggctgat      1833 
Trp 

agcccacccc gcccccactg cagctcccac aaagcactct aagggagatg gggctgagga   1893 

cagctcaaaa aggagaggac aggcctgcag ggcccacttg cccagcacca aggcgagctc   1953 

cgggctcaga ccaactccaa ggtcagctgg ccacagccca ggcctcctct gaagcaggga   2013 

ccagagggag ccaggcagct gtgtacagtg agaagggatc cggatggggg agctctgtac   2073 

agagggctgg tgattgtaaa aatttctttt gtaaagtaga agttgggggt ggggtgggtg   2133 

ctggctgcaa aaatttctgg cttctcttac ccctattgcc cccggcaata aattgtttct   2193 

atatgccaaa aaaaaaaaaa aa                                            2215 

 
           
             10  
             513  
             PRT  
             Homo sapiens  
           
            10 

Met Ala Glu Val Gly Glu Ile Ile Glu Gly Cys Arg Leu Pro Val Leu 
  1               5                  10                  15 

Arg Arg Asn Gln Asp Asn Glu Asp Glu Trp Pro Leu Ala Glu Ile Leu 
             20                  25                  30 

Ser Val Lys Asp Ile Ser Gly Arg Lys Leu Phe Tyr Val His Tyr Ile 
         35                  40                  45 

Asp Phe Asn Lys Arg Leu Asp Glu Trp Val Thr His Glu Arg Leu Asp 
     50                  55                  60 

Leu Lys Lys Ile Gln Phe Pro Lys Lys Glu Ala Lys Thr Pro Thr Lys 
 65                  70                  75                  80 

Asn Gly Leu Pro Gly Ser Arg Pro Gly Ser Pro Glu Arg Glu Val Pro 
                 85                  90                  95 

Ala Ser Ala Gln Ala Ser Gly Lys Thr Leu Pro Ile Pro Val Gln Ile 
            100                 105                 110 

Thr Leu Arg Phe Asn Leu Pro Lys Glu Arg Glu Ala Ile Pro Gly Gly 
        115                 120                 125 

Glu Pro Asp Gln Pro Leu Ser Ser Ser Ser Cys Leu Gln Pro Asn His 
    130                 135                 140 

Arg Ser Thr Lys Arg Lys Val Glu Val Val Ser Pro Ala Thr Pro Val 
145                 150                 155                 160 

Pro Ser Glu Thr Ala Pro Ala Ser Val Phe Pro Gln Asn Gly Ala Ala 
                165                 170                 175 

Arg Arg Ala Val Ala Ala Gln Pro Gly Arg Lys Arg Lys Ser Asn Cys 
            180                 185                 190 

Leu Gly Thr Asp Glu Asp Ser Gln Asp Ser Ser Asp Gly Ile Pro Ser 
        195                 200                 205 

Ala Pro Arg Met Thr Gly Ser Leu Val Ser Asp Arg Ser His Asp Asp 
    210                 215                 220 

Ile Val Thr Arg Met Lys Asn Ile Glu Cys Ile Glu Leu Gly Arg His 
225                 230                 235                 240 

Arg Leu Lys Pro Trp Tyr Phe Ser Pro Tyr Pro Gln Glu Leu Thr Thr 
                245                 250                 255 

Leu Pro Val Leu Tyr Leu Cys Glu Phe Cys Leu Lys Tyr Gly Arg Ser 
            260                 265                 270 

Leu Lys Cys Leu Gln Arg His Leu Thr Lys Cys Asp Leu Arg His Pro 
        275                 280                 285 

Pro Gly Asn Glu Ile Tyr Arg Lys Gly Thr Ile Ser Phe Phe Glu Ile 
    290                 295                 300 

Asp Gly Arg Lys Asn Lys Ser Tyr Ser Gln Asn Leu Cys Leu Leu Ala 
305                 310                 315                 320 

Lys Cys Phe Leu Asp His Lys Thr Leu Tyr Tyr Asp Thr Asp Pro Phe 
                325                 330                 335 

Leu Phe Tyr Val Met Thr Glu Tyr Asp Cys Lys Gly Phe His Ile Val 
            340                 345                 350 

Gly Tyr Phe Ser Lys Glu Lys Glu Ser Thr Glu Asp Tyr Asn Val Ala 
        355                 360                 365 

Cys Ile Leu Thr Leu Pro Pro Tyr Gln Arg Arg Gly Tyr Arg Lys Leu 
    370                 375                 380 

Leu Ile Glu Phe Ser Tyr Glu Leu Ser Lys Val Glu Gly Lys Thr Gly 
385                 390                 395                 400 

Thr Pro Glu Lys Pro Leu Ser Asp Leu Gly Leu Leu Ser Tyr Arg Ser 
                405                 410                 415 

Tyr Trp Ser Gln Thr Ile Leu Glu Ile Leu Met Gly Leu Lys Ser Glu 
            420                 425                 430 

Ser Gly Glu Arg Pro Gln Ile Thr Ile Asn Glu Ile Ser Glu Ile Thr 
        435                 440                 445 

Ser Ile Lys Lys Glu Asp Val Ile Ser Thr Leu Gln Tyr Leu Asn Leu 
    450                 455                 460 

Ile Asn Tyr Tyr Lys Gly Gln Tyr Ile Leu Thr Leu Ser Glu Asp Ile 
465                 470                 475                 480 

Val Asp Gly His Glu Arg Ala Met Leu Lys Arg Leu Leu Arg Ile Asp 
                485                 490                 495 

Ser Lys Cys Leu His Phe Thr Pro Lys Asp Trp Ser Lys Arg Gly Lys 
            500                 505                 510 

Trp