TRANSCRIPTION FACTOR-SELECTIVE NUCLEAR TRANSPORT RECEPTORS REQUIRED FOR IMMUNE RESPONSE ACTIVATION

Abstract: Biological molecules are herein identified and characterized as specific transport receptors which control the entry of NF- kappa B and related transcription factors into the cell nucleus for activation of the immune response, <i>inter alia</i>. Methods are provided which employ the transcription factor-nuclear transport receptor sequences to identify compounds that modulate biological and/or pharmacological activity of the receptors and hence regulate cell and tissue physiology.

Description: TRANSCRTPTION FACTOR-SELECTIVE NUCLEAR TRANSPORT RECEPTORS REQUIRED FOR IMMUNE RESPONSE ACTIVATION Field of the Invention The present invention relates to nucleic acid and amino acid sequences pertaining to nuclear receptors which selectively facilitate the entry of transcription factors into the cell nucleus and thereby regulate gene expression and cell/tissue physiology. The nuclear import biomolecules are specifically required for the import of Nuclear Factor-kappaB (NF-κB) and related transcription activators required for the immune response and the mediation of related pathophysiological disorders. Molecular sequences are provided for the design and synthesis of entities that modulate biological and/or pharmacological activity of native nuclear receptors. The sequences are also provided for employment to identify small molecule compounds that modulate biological and/or pharmacological activity of the native receptors. The invention is also drawn toward the study, prevention, diagnosis, and treatment of pathophysiological disorders related to the family of nuclear receptors. Background of the Invention Developmental processes and physiological responses rely on signal transduction pathways that culminate with the selective nuclear entry of transcription factors and other regulatory proteins to control gene expression. See, e.g., Nigg, E. A., Nucleocytoplasmic Transport: Signals, Mechanisms and Regulation, Nature, 386:119 (1997); Mattaj, I. W., et al., L. Nucleocytoplasmic Transport: The Soluble Phase, Annu. Rev. Biochem., 67:265 (1998). Protein import commences in the cytoplasm by the interaction between the protein that is targeted to the nucleus and its transport receptor via the nuclear localization signal (NLS), a short stretch of basic amino acids responsible for nuclear targeting. The transport receptor docks the complex to the nuclear pore and escorts the cargo through the pore. The difference between the various transport pathways is the use of distinct transport receptors. The activation of an immune response relies to a large extent on the induced translocation of proteins from the cytoplasm into the nucleus. The Nuclear Factor- kappaB proteins (NF-κB), for example are involved in both Drosophila and mammalian responses to infection, and are held in the cytoplasm by interaction with an inhibitor protein, IκB. Upon signaling the IκB protein becomes phosphorylated and degraded, and the NF-κB protein is then free to move into the nucleus. See, e.g., Kopp, E. B., et al., Curr. Opin. Immunol., 11: 13 (1999). In Drosophila three NF-κB proteins have been identified, dorsal (Steward, R., Dorsal, An Emryonic Polarity gene in Drosophila, is Homologous to the Vertrbrate Proto-Oncogene, c-rel, Science 238, 692-694(1987)), Dif (Ip, Y. T., et al., Dif, A Dorsal-Related Gene that Mediates an Immune Response in Drosophila, Cell 75, 753-763.(1997)), and Relish (Dushay, M. S., etal, Origins of Immunity: Relish, a Compound Rel-Like Gene in the Antibacterial Defense of Drosophila, Proc. Natl. Acad. Sci. USA, 93: 10343 (1996)). The two former are retained in the cytoplasm by interaction with the IκB analogous cactus protein in Drosophila. Upon an immune response, these proteins are translocated into the nucleus. Kopp, E. B., et al., Curr. Opin. Immunol., 11:13 (1999). The NF-κB proteins are moreover key mammalian transcription factors which regulate, for example, the expression of pro- inflammatory genes. Despite the well defined signaling pathways that lead to activation of the NF-κB proteins, little has previously been known about the discriminating mechanism, namely the transport receptor, which facilitates selective translocation of NF-κB and related transcrimption factors into the nucleus. The identifiction of a family of transport receptors which control the entry of NF- kB and related transcription factors into the cell nucleus, including mammalian cells, would convey an ability to specifically regulate gene expression and thereby facilitate broad therapeutic potential in the control of a myriad of pathophysiological disorders including but not limited to inflammation, asthma, rheumatoid arthritis (RA), chronic obstructive pulmonary disease (COPD), angiogenesis, septic shock, lung fibrosis, glomerulonephritis, artherosclerosis, AIDS, and apoptotic disorders. Summary of the Invention The present invention is directed to an isolated and purified polynucleotide molecule comprising a nucleic acid sequence which encodes a polypeptide having at least about 90% homology to a member selected from (SEQ ID NO:3, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ JD NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ED NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725). Isolated and purified polynucleotides of the present invention include but are not limited to sequences which comprise SEQ ID NO: 1 and/or SEQ ID NO:2. The current invention is directed to a purified polypeptide selected from (SEQ ID NO:3, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO.4 positions 575-725, SEQ ID NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ED NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725) or a variant thereof as defined herein. The instant invention is further directed to methods of identifying compounds that modulate a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor, comprising: (a) combining a candidate compound modulator with a polypeptide comprising a sequence having at least about 90% homology to a member selected from (SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575- 725, SEQ ID NO:5, SEQ ID NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725), and (b) measuring an effect of the candidate compound modulator on the biological and/or pharmacological activity of the polypeptide. Detailed Description of the Invention Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this invention belongs. All publications and patents referred to herein are incorporated by reference. Biological activity as used herein in reference to a transcription factor-selective nuclear transport receptor polypeptide, e.g., SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5, SEQ ID NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ID NO:5 positions 50- 350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725, refers to the ability of the peptide to interact with and/or bind to the rel or NF-κB family of proteins, including but not limited to NF-κB, the NF-κB NLS motif, members of the rel protein family, dorsal (GENBANK access M23702), Dif, relish, DPSE-1 (GENBANK accession AF132561) and the Human homolog O00410, shaq proteins including but not limited to the product of human oncogene CAN 214 (CAN/Nup214) and Human homolog NM_005085.1, human CRMl Y08614, STATs (e.g., STAT-1 (P42224), STAT-2 (NM005419), STAT-4 (L78440), STAT-5 (L41142), STAT-5B (U47686), and STAT-6 (NM003153), SMADs (e.g., Smad 6 (AFO35528 and AAB 94137), and Smad 3 (BAA22032 ), pRELB (GENBANK access NP 006500), KBF2 Human (GENBANK access Q00653) (NFkappa B pi 00), AAB21124 (NFkappa b p50 homolog), A42024 (NFkappa b pl00), A37867 (NFkappa b 50k precursor), AAB23437 (NFkappa b pl05 homolog), CAA8052 (NFkappa b ρ65), pRAI NP_006654, pNFKBIE NP_004547, KBFl human (GENBANK access P19838) (NFkappa b pl05), CAA87683 (NFkappa b), RELB iuman (GENBANK access Q01201 ) (RELB/I-REL), BCL3_human (GENBANK access P20749), AAA36408 (NFkappa b), and AAA36127 (I-REL). These are biological molecules whose nuclear entry is expected to be regulated by transcription factor-selective nuclear transport receptor polypeptides described herein. They are all implicated in the activation of the inflammatory response. Pharmacological activity as used herein in reference to a transcription factor- selective nuclear transport receptor polypeptide, refers to the ability of the biomolecule to mediate cell processes related to the immune response and inflammation including but not limited to inflammation, asthma, rheumatoid arthritis (RA), chronic obstructive pulmonary disease (COPD), angiogenesis, septic shock, lung fibrosis, glomerulonephritis, artherosclerosis, AIDS, and apoptotic disorders; and/or the direct or indirect transcriptional activation of one or more genes. See, e.g., Chen F., et al., Clin. Chem., 45(1):7 (1999). As depicted as used herein refers the sequence as well as inherent variants thereof, e.g., functional derivatives that demonstrate or perform substantially the same biological and/or pharmacological activity in substantially the same way. 'As depicted' is therefore intended to encompass variants of the subject molecule contemplated herein. Variant as used herein refers to sequences substantially as shown having changes, e.g., a polypeptide sequence comprising a sequence which differs from the sequence referred to by at least one amino acid substitution, addition, or deletion, preferrably a conservative amino acid substitution, that demonstrate or perform substantially the same biological and/or pharmacological activity in substantially the same way, as well as truncated versions of these variants. However, variant as used herein is intended to encompass all biologically effective dominant negative mutants. The term modulation is used herein to refer to the capacity to either enhance or inhibit a function of a biological molecule including, but not limited to, a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor, or to the capacity to either enhance or inhibit a functional property of a nucleic acid regulatory or coding region. Modulate physiology as used herein refers to the biophysiological regulation of cells and/or tissue and the treatment of pathophysiological disorders related thereto. Direct administration as used herein refers to the direct administration of nucleic acid molecules, peptides, or compounds as well as contemplated derivatives/variants of the present invention. Direct administration includes but is not limited to ex vivo as well as in vivo gene therapy techniques. Purified as used herein refers to molecules, either nucleic acid or amino acid sequences, that are removed from their natural environment and isolated or separated from at least one other component with which they are naturally associated. Expression vector as used herein refers to nucleic acid vector constructions to direct the transcription of nucleic acid regions in host cells. Expression vectors include but are not limited to plasmids, retroviral vectors, viral and synthetic vectors. Transformed host cells as used herein refer to cells which harbor one or more nucleic acids of the present invention. Transcription Factor-Selective Nuclear Transport Receptors Members Only (mbo) (SEQ ID NO: 1), a Drosophila gene described herein, encodes a nuclear pore receptor (SEQ ID NO:3) that mediates import of dorsal NF- B at the level of the nuclear pore and is required for activation of the Drosophila humoral immune response. The mbo gene product (SEQ ED NO:3) localizes to the nuclear envelope and is homologous to mammalian proteins including human Nup88 (SEQ ID NO:4) as well as rat Nup88 (SEQ ID NO: 5) (also known as Nup84). Nup88 is reported to localize to the cytoplasmic filaments of the nuclear pores to provide binding sites for nuclear import substrates; however, the function of mammalian Nup88 has previously been unknown. Fornerod, M, et al., EMBO J. 16:807 (1997); Bastos, R., et al., J. Cell Biol. 137:989 (1997). The previously reported mammalian Nup88 molecules are herein identified and characterized as specific transport receptors which control the entry of NF- KB and related transcription factors into the cell nucleus Mbo (SEQ ID NO:3) demonstrates extensive homology to Nup88 (SEQ ID NO:4) especially considering the evolutionary divergence of the species. Overall the amino acid position identity is 24% and the homology is 52% and extends throughout the length of each. The most striking feature of both Nup88 and Mbo is their C-terminal regions which have a high probability score of forming a coiled coil structure. Such structures are often found to mediate protein interactions. This region of human Nup88 is important for its association with the nuclear pore complex. Bastos, R., et al., J. Cell Biol. 137:989 (1997). The proteins also share enhanced sequence similarity within their N-terminal and central domains. Moreover, when human Nup88 is overexpressed in tissue-culture cells, the protein becomes distributed throughout the cytoplasm, but not in the nucleus. Similarly, in Drosophila embryos which overexpresses Mbo in tracheal cells using the UAS-GAL4 system, staining for Mbo reveals a uniform cytoplasmic distribution which demonstrates that mbo (SEQ ID NO:3) is normally localized on the cytoplasmic side of the nuclear envelope. The immune response is an example of a process which to a large extent relies on the induced translocation of proteins from the cytoplasm into the cell nucleus. NF-kB transcription factors are involved Drosophila as well as mammalian responses to infection and are maintained in the cytoplasm. In addition to its role in the immune response, Mbo is dynamically expressed and required during several distinct developmental processes. During tracheal branch fusion, for example, Mbo appears to be involved in the morphogenetic process giving rise to a bicellular anastomosis connecting two independent branches. The nuclear translocation of the Drosophila NF-κB protein dorsal is induced to determine the dorso-ventral polarity of the early embryo. Morisato, O., etal., Ann. Rev. Gen. 29:371 (1995). Dorsal translocation is also part of the activation of the larval immune response to infection following the activation of the same signaling cascade. Lemaitre, B., et al, EMBO J., 14:536 (1995). Moreover, the Drosophila homologue of mammalian STATs is striking similar in its zygotic expression pattern and tracheal mutant phenotype to Mbo. STAT is also involved in the immune response of vertebrates, and is expected to have a similar role in Drosophila. Barillas-Mury, C, et al., EMBO J., 18:959 (1999). Thus, STATs appear, in addition to NF-κBs, to be targets for the family of transcription factor-selective nuclear transport receptors decribed herein (e.g., SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5). The subcellular localization of dorsal in homozygous Mbo larvae and their heterozygous siblings was investigated and is herein disclosed. Both mutant and heterozygous larvae where reared together, and in non-challenged larvae of both genotypes dorsal is demonstrated to be predominantly cytoplasmic. When the larvae were infected with a needle dipped in a bacterial culture, dorsal translocated into the nucleus within 45 minutes in the heterozygous larvae. Mbo mutant larvae that received the same treatment demonstrated no nuclear localization of dorsal. It is also demonstrated that dorsal becomes translocated in the lymph gland tissues in wild type larvae upon infection. Nuclear dorsal in lymph glands is not detected in unchallenged larvae; after challenge all larvae tested (n=100) display nuclear localization of dorsal in the lymph glands. The lymph glands of mutant larvae, however, show dorsal mainly in the cytoplasm after induction. Wild type larvae nuclear translocation of the rel proteins dorsal and Dif is followed by the rapid transcriptional activation of genes encoding antimicrobial peptides. Petersen, U. M., et al., EMBO J., 14:3146 (1995). A reporter construct of the inducible Cecropin A promoter coupled to LacZ (cec-lacZ) is strongly induced in wild type larvae upon infection; whereas it is non responsive in Mbo mutants. Further analysis of the inducible expression of the genes for antimicrobial peptides Drosomycin and Diptericin by Northern blot hybridizations revealed that their induction is severely impaired in Mbo mutant larvae. Mbo (SEQ ID NO:3) is required for the inducible translocation of the Drosophila NF-κB homologue dorsal into the nucleus and the concomitant transcriptional activation of the genes coding for antimicrobial peptides. Immunoprecipitations were carried out, using a polyclonal antiserum that has been shown before to immunoprecipitate dorsal from protein extracts, to demonstrate that Mbo directly interacts with dorsal. Gillespie, S. H., et al., Mol. Cell Biol., 14:3559 (1994). It is moreover demonstrated that Mbo is not required for general protein import and/or for mRNA export. Furthermore, de novo protein production can occur in Mbo mutants which demonstrates a fully functional basal transcription and translation machinery. Because the basic transport machinery across the nuclear pore is functional in Mbo mutants and since in situ antibody stainings with anti Mbo antiserum localize the protein in a punctuated ring around the nuclear envelope Mbo appears unequivocally to directly participate in the nuclear translocation of dorsal. Regulated nuclear import of the Drosophila NF-kB protein dorsal that normally occurs upon a immune response is prevented in Mbo mutants, despite the fact that upstream signaling events resulting in the degradation of the IkB protein cactus are intact. Moreover, as a result, downstream target genes which encode antimicrobial peptides are not activated. Mbo mutants do not show any defects in RNA export, classical nuclear localization signal (NLS)-dependent import or import of phosphorylated MAP kinase and cyclin B into the nucleus. Immunoprecipitations from embryonic extracts show complex formation between Mbo and dorsal. Pharmacological Significance The identification of Mbo (SEQ ED NO:3) specifically required for the nuclear import of the Drosophila NF-kB protein dorsal and the corollary immune response, as well as the characterization of the pharmacological significance of mammlian homologues (SEQ ID NO:4, SEQ ED NO: 5) presents valuable opportunities to control the nuclear import and therefore physiological effects of NF-kB (e.g., modulation of immune response) and related transcription factors. Human Shag, for example, was originally isolated as an oncogene where two different translocations affect the human gene and cause either a subtype of acute myeloid leukemia or acute undifferentiated leukemia. The mouse knockout of Shaq is lethal in utero and phenotypes include defects in nuclear RNA export whereas classical NLS- dependent import is not affected. A two hundred amino acid region in the coil domain of the Drosophila homologue of Shaq is identified as necessary for binding to the N-terminal 200 aa of >o (SEQ ID NO:3). It is the prediction that drugs targeting the interaction of these domains in mammalian Mbo homologues (e.g., SEQ ID NO:4, SEQ DD NO:5) will inhibit translocation of Rel-proteins. Data also indicate that certain STAT factors require Mbo for their translocation. The transcription factor-selective nuclear transport receptors, particularly mammalian Mbo homologues and variations contemplated herein, provide new targets for drugs to specifically regulate gene expression and thereby facilitates broad therapeutic potential in the control of a myriad of pathophysiological disorders including but not limited to inflammation, asthma, rheumatoid arthritis (RA), chronic obstructive pulmonary disease (COPD), angiogenesis, septic shock, lung fibrosis, glomerulonephritis, artherosclerosis, AIDS, and apoptotic disorders. Accordingly, the ability to modulate a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor polypeptide is expected to confer the ability to ameliorate these pathophysiological conditions. The human homologue (SEQ ID NO:4) and other homologues (e.g., SEQ ID NO:5) of the nuclear import protein Mbo in Drosophila (SEQ ID NO:3) are valuable targets for selective drug intervention of transcription factor, e.g., NF-kB, entry into the nucleus. The present invention therefore relates to methods of identifying molecules capable of modulating a biological and/or pharmacological activity of a transcription factor- selective nuclear transport receptor polypeptide, e.g., SEQ ED NO:3, SEQ ID NO:4, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5, SEQ ID NO:5 positions 1-200, SEQ ID NO: 5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575- 725. Polynucleotide sequences which encode these molecules and variants thereof contemplated herein are particularly preferred embodiment of the present invention. Variants The present invention relates to variants of nucleic acid sequences sequences and amino acid sequences substantially as shown or known, which have changes, e.g., a polypeptide sequence comprising a sequence which differs from the sequence referred to by at least one amino acid substitution, preferrably a conservative amino acid substitution (or a nucleic acid sequence which encodes therefor), that demonstrate or perform substantially the same biological and/or pharmacological activity in substantially the same way, as well as molecules which comprise truncated versions. However, variant as used herein is intended to encompass biologically effective dominant negative mutants. Preferred variants of molecules described herein, for example, comprise an amino acid sequences having at least 90% amino acid sequence homology (identity) to a sequence selected from SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5, SEQ ID NO:5 positions 1- 200, SEQ ED NO:5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ ED NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725 or a biologically and/or pharmacologically active substantial fragment thereof. A variant of a transcription factor-selective nuclear transport receptor polypeptide may have an amino acid sequence that is different by one or more amino acid substitutions. Embodiments which comprise amino acid deletions and/or additions are also contemplated. The variant may have conservative changes (amino acid similarity), wherein a substituted amino acid has similar structural or chemical properties, for example, the replacement of leucine with isoleucine. A variant may have nonconservative changes, e.g., replacement of a glycine with a tryptophan. Guidance in determining which and how many amino acid residues may be substituted, inserted or deleted without abolishing biological or proposed pharmacological activity may be reasonably inferred in view of this disclosure and may be further be found using computer programs well known in the art, for example, DNAStar software. Amino acid substitutions may be made, for instance, on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues as long as a biological and/or pharmacological activity of the native molecule is retained. Negatively charged amino acids, for example, include aspartic acid and glutamic acid; positively charged amino acids include lysine and arginine; and amino acids with uncharged polar head groups having similar hydrophilicity values include leucine, isoleucine, valine; glycine, alanine; asparagine, glutamine; serine, threonine phenylalanine, and tyrosine. However, in the construction of biologically effective dominant negative mutants at least one amino acid residue position at an active site required for biological and/or pharmacological activity in the native peptide is changed to produce an agent or entity having reduced activity or which is devoid of detectable activity. Suitable substitutions of amino acids include the use of a chemically derivatized residue in place of a non-derivatized residue. D-isomers as well as other known derivatives may also be substituted for the naturally occurring amino acids. See, e.g., U.S. Patent No. 5,652,369, Amino Acid Derivatives, issued July 29, 1997. Example substitutions are set forth in TABLE 1 as follows: Table 1 "Homology" as used in this description is a measure of the identity of nucleotide sequences or amino acid sequences. In order to characterize the homology, subject sequences are aligned so that the highest order homology (match) is obtained. "Identity" per se has an art-recognized meaning and can be calculated using published techniques. Computer program methods to determine identity between two sequences, for example, include DNAStar software (DNAStar Inc., Madison, WI); the GCG program package (Devereux, J., et al, Nucleic Acids Research (1984) 12(1):387); BLASTP, BLASTN, FASTA (Atschul, S. F. et al., J Molec Biol (1990) 215:403). Homology (identity) as defined herein is determined conventionally using the well known computer program, BESTFIT (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wis., 53711). When using BESTFIT or any other sequence alignment program to determine whether a particular sequence is, for example, about 90% homologous to a reference sequence, according to the present invention, the parameters are set such that the percentage of identity is calculated over the full length of the reference nucleotide sequence or amino acid sequence and that gaps in homology of up to about 10% of the total number of nucleotides in the reference sequence are allowed. Eighty percent of homology is therefore determined, for example, using the BESTFIT program with parameters set such that the percentage of identity is calculated over the full length of the reference sequence, e.g., SEQ ID NO:3, and gaps of up to 10% of the total number of amino acids in the reference sequence are allowed, and wherein up to 10% of the amino acid residues in the reference sequence may be deleted or substituted with another amino acid, or a number of amino acids up to 10% of the total amino acid residues in the reference sequence may be inserted into the reference sequence. Percent homologies are likewise determined, for example, to identify preferred species, within the scope of the claims appended hereto, which reside within the range of about 90 percent to 100 percent homology to SEQ ID NO: 3 as well as biologically and/or pharmacologically active derivatives thereof. Percentage similarity (conservative substitutions) between two polypeptides may also be scored by comparing the amino acid sequences of the two polypeptides by using programs well known in the art, including the BESTFIT program, by employing default settings for determining similarity. Transcription factor-selective nuclear transport receptor molecules described herein, e.g., SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ED NO:4 positions 50-350, SEQ ID NO:4 positions 550- 740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5, SEQ ID NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725, as well as nucleic acid molecules which comprise a sequence which encodes therefor, are used in screening assays to identify antagonists or inhibitors which bind to, or interact with the nuclear receptor molecules , emulate its substrate, or otherwise inactivate one of the biomolecule or compete biologically, e.g., competitive interaction or competitive binding inhibition. The nuclear receptor molecules are also used in screening assays to identify agonists which agonize or mimic the biological and/or pharmacological activity, induce the production of or prolong the biological halflife of the molecule in vivo or in vitro. Generally Acceptable Vectors In accordance with the present invention, polynucleotide sequences which encode a nuclear transport receptor described herein or a corresponding polypeptide, dominant negative mutant versions, fusion proteins, or antisense molecules may be used in recombinant DNA molecules that direct the expression of the respective molecule in appropriate host cells. Due to the inherent degeneracy of the genetic code, other DNA sequences which encode substantially the same or a functionally equivalent amino acid sequence may be used to clone and express the transcription factor-selective nuclear transport receptor polypeptide, as well as variations thereto and dominant negative mutants thereof. As will be understood by those of skill in the art, it may be advantageous to produce nucleotide sequences possessing non-naturally occurring codons. SEQ ID NO:2, for example, may be recombinantly expressed by molecular cloning into an expression vector containing a suitable promoter and other appropriate transcription regulatory elements, and transferred into prokaryotic or eukaryotic host cells to produce the native biomolecule. Techniques for such manipulations are fully described in Sambrook, J., et al., Molecular Cloning Second Edition, Cold Spring Harbor Press (1990), and are well known in the art. Expression vectors are described herein as nucleic acid sequences for the transcription of embodiments of the present invention. Such vectors can be used to express nucleic acid sequences in a variety of hosts such as bacteria, bluegreen algae, plant cells, insect cells, fungal cells, human, and animal cells. Specifically designed vectors allow the shuttling of DNA between hosts such as bacteria-yeast, or bacteria-animal cells, or bacteria-fungal cells, or bacteria-invertebrate cells. A variety of mammalian expression vectors may be used to express nuclear transport receptor molecules as well as variants and derivatives contemplated herein. Commercially available mammalian expression vectors which are suitable for recombinant expression, include but are not limited to, pcDNA3 (Invitrogen), pMClneo (Stratagene), pXTl (Stratagene), pSG5 (Stratagene), EBO-pSV2-neo (ATCC 37593) pBPV-l(8-2) (ATCC 37110), pdBPV-MMTneo(342-12) (ATCC 37224), pRSVgpt (ATCC 37199), pRSVneo (ATCC 37198), pSV2-dhfr (ATCC 37146), pUCTag (ATCC 37460), and 1ZD35 (ATCC 37565), pLXIN and pSIR (CLONTECH), pIRES-EGFP (CLONTECH). INVITROGEN corporation provides a wide variety of commercially available mammalian expression vector/systems which can be effectively used with the present invention. INVITROGEN, Carlsbad, CA. See, also, PHARMINGEN products, vectors and systems, San Diego, CA. Baculoviral expression systems may also be used with the present invention to produce high yields of biologically acive transcription factor-selective nuclear transport receptor. Vectors such as the CLONETECH, BacPak™ Baculovirus expression system and protocols are preferred which are commercially available. CLONTECH, Palo Alto, CA. Miller, L.K., et al, Curr. Op. Genet. Dev. 3:97 (1993); O'Reilly, D.R., et al, Baculovirus Expression Vectors: A Laboratory Manual, 127. Vectors such as the INVITROGEN, MaxBac™ Baculovirus expression system, insect cells, and protocols are also preferred which are commercially available. INVITROGEN, Carlsbad, CA. Example Host Cells Host cells transformed with a nucleotide sequence which encodes the nuclear transport receptor mlecules described herein may be cultured under conditions suitable for the expression and recovery of the encoded protein from cell culture. Preferred embodiments for use in methods of the invention are host cells transformed with a purified polynucleotide comprising a nucleic acid sequence which encodes a polypeptide comprising a sequence as depicted in SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5, SEQ ID NO: 5 positions 1-200, SEQ ID NO: 5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and/or SEQ ID NO:5 positions 575-725 or a contemplated homolog or variant thereof. Cells of this type or preparations made from them may be used to screen for modulators of the biological and/or pharmacological activity of the recited molecules. Eukaryotic recombinant host cells are especially preferred. Examples include but are not limited to yeast, mammalian cells including but not limited to cell lines of human, bovine, porcine, monkey and rodent origin, and insect cells including but not limited to Drosophila and silkworm derived cell lines. Cell lines derived from mammalian species which may be suitable and which are commercially available, include but are not limited to, L cells L-M(TK-) (ATCC CCL 1.3), L cells L-M (ATCC CCL 1.2), 293 (ATCC CRL 1573), Raji (ATCC CCL 86), CV-1 (ATCC CCL 70), COS-1 (ATCC CRL 1650), COS-7 (ATCC CRL 1651), CHO-K1 (ATCC CCL 61), 3T3 (ATCC CCL 92), NIH/3T3 (ATCC CRL 1658), HeLa (ATCC CCL 2), C127I (ATCC CRL 1616),BS-C-1 (ATCC CCL 26) and MRC-5 (ATCC CCL 171). The expression vector may be introduced into host cells to express a transcription factor-selective nuclear transport receptor polypeptide and derivatives thereof via any one of a number of techniques including but not limited to transformation, transfection, lipofection, protoplast fusion, and electroporation. Commercially available kits applicable for use with the present invention for hererologous expression, including well- characterized vectors, transfection reagents and conditions, and cell culture materials are well-established and readily available. CLONTECH, Palo Alto, CA; INVITROGEN, Carlsbad, CA; PHARMINGEN, San Diego, CA; STRATAGENE, LaJolla, CA. The expression vector-containing cells are clonally propagated and individually analyzed to determine the level of transcription factor-selective nuclear transport receptor polypeptide production. Identification of host cell clones which express a transcription factor-selective nuclear transport receptor polypeptide may be performed by several means, including but not limited to immunological reactivity with antibodies described herein, and/or the presence of host cell-associated specific biological activity, and/or the ability to covalently cross-link specific substrate to a transcription factor-selective nuclear transport receptor polypeptide with the bifunctional cross-linking reagent disuccinimidyl suberate or similar cross-linking reagents. Transcription factor-selective nuclear transport receptor polypeptides may be expressed to facilitate practicing methods of the invention as a recombinant protein with one or more additional polypeptide domains added to facilitate protein purification. Such purification facilitating domains include, but are not limited to, metal chelating peptides such as histidine-tryptophan modules that allow purification on immobilized metals (Porath, J., Protein Exp. Purif. 3:263 (1992)), protein A domains that allow purification on immobilized immunoglobulin, and the domain utilized in the FLAGS extension/affinity purification system (Immunex Corp, Seattle WA). The inclusion of a cleavable linker sequences such as Factor XA or enterokinase (Invitrogen, San Diego CA) between the purification domain and the coding region is useful to facilitate purification. Systems such as the CLONTECH, TALON™ nondenaturing protein purification kit for purifying 6xHis-tagged proteins under native conditions and protocols are preferred which are commercially available. CLONTECH, Palo Alto, CA. In addition, a host cell strain may be chosen for its ability to modulate the expression of the inserted sequences or to process the expressed protein in the desired fashion. Such modifications of the polypeptide include, but are not limited to, acetylation, carboxylation, glycosylation, phosphorylation, lipidation and acylation. Post-translational processing which cleaves a nascent form of the protein may also be important for correct insertion, folding and/or function. Different host cells such as CHO, HeLa, MDCK, 293, WI38, NIH-3T3, HEK293 etc., have specific cellular machinery and characteristic mechanisms for such post-translational activities and may be chosen to ensure the correct modification and processing of the introduced, foreign protein. For long-term, high-yield production of recombinant proteins, stable expression is preferred. For example, cell lines which stably express SEQ ID NO:4, for example, may be transformed using expression vectors which contain viral origins of replication or endogenous expression elements and a selectable marker gene. Following the introduction of the vector, cells may be allowed to grow for 1-2 days in an enriched media before they are switched to selective media. The purpose of the selectable marker is to confer resistance to selection, and its presence allows growth and recovery of cells which successfully express the introduced sequences. Resistant clumps of stably transformed cells can be proliferated using tissue culture techniques appropriate to the cell type. The nuclear transport receptor peptides can be produced in the yeast S.cerevisiae following the insertion of the optimal cDNA cistron into expression vectors designed to direct the intracellular or extracellular expression of the heterologous protein. In the case of intracellular expression, vectors such as EmBLyex4 or the like are ligated to the beta subunit cistron. See, e.g., Rinas, U., et al, Biotechnology, 8:543 (1990); Horowitz, B., et al, J. Biol. Chem., 265:4189 (1989). For extracellular expression, the receptor 'cistron' is ligated into yeast expression vectors which may employ any of a series of well- characterized secretion signals. A variety of protocols for detecting and measuring the expression of the novel molecule as well as functional derivatives thereof, using either polyclonal or monoclonal antibodies specific for the protein are known in the art. See Example I. Examples include enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA) and fluorescent activated cell sorting (FACS). A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering epitopes may be employed. Well known competitive binding techniques may also be employed. See, e.g., Hampton, R, et al. (1990), Serological Methods - a Laboratory Manual, APS Press, St Paul Minn.; Maddox, D.E., et al, J. Exp. Med. 158:1211. Example Embodiments A purified polynucleotide is preferred which comprises a nucleic acid sequence which encodes a polypeptide having at least about 90% homology to a member selected from (SEQ ED NO:3, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ED NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ ED NO:5 positions 550-740, and SEQ ED NO:5 positions 575-725). Methods are provided to screen compounds individually, or libraries of compounds, for the identification of compounds which have the ability to modulate a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor polypeptide, particularly SEQ ID NO:4 and/or SEQ ID NO:5. The present invention is also directed to methods of screening for compounds which modulate the expression (transcription and/or translation) of DNA or RNA which encode transcription factor-selective nuclear transport receptors. Compounds which modulate these activities may be DNA, RNA, peptides, proteins, or non-proteinaceous organic molecules (e.g., small molecule compounds). Example assays described herein are used to identify agents which modulate nuclear transport receptor biological interaction with NF-κB family peptides and/or a pharmacological activity of a transcription factor-selective nuclear transport receptor polypeptide. Such modulating agents are contemplated for therapeutic purposes, i.e., administration for the treatment of pathophysiological disorders related to the biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor, e.g., SEQ ID NO:4. Compounds may modulate an ultimate biological and/or pharmacological activity by increasing or attenuating the expression of DNA or RNA encoding a transcription factor-selective nuclear transport receptor. Compounds that modulate the expression of DNA or RNA encoding such a receptor or the function of the polypeptide may be detected by a variety of assays. The assay may be a simple "yes/no" assay to determine whether there is a change in expression or activity. The assay may be made quantitative by comparing the expression or function of a test sample with the levels of expression or function in a standard sample. The transcription factor-selective nuclear transport receptor polypeptides as depicted as well as contemplated variants can be used for screening therapeutic compounds in any of a variety of drug screening techniques. The fragment or entity employed in such a test may be free in solution, affixed to a solid support, borne on a cell surface, or located intracellularly. The abolition or modulation of activity or the formation of binding complexes, between a nuclear transport receptor molecule and the agent being tested, may be measured, for example, by means provided (see, Examples appended hereto). Accordingly, the present invention provides a method for screening a plurality of compounds for specific binding affinity with a polypeptide, e.g., SEQ ED NO:4, or a variant thereof contemplated herein, comprising providing a plurality of compounds; combining an example nuclear transport receptor polypeptide with each of a plurality of compounds for a time sufficient to allow binding under suitable conditions; and detecting binding of an embodiment of a nuclear transport receptor, to each of the plurality of compounds, thereby identifying the compounds which specifically bind the transport receptor polypeptide. Disruption of SEQ ID NO:4 binding to NF-κB, for example, is readily measurable by means of competitive binding assays well known in the art. Methods of identifying compounds that modulate a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor polypeptide, e.g., SEQ ID NO:4, are generally preferred, which comprise combining a candidate compound modulator with a purified polypeptide comprising the amino acid sequence as depicted in SEQ ED NO:4 or a variant of SEQ ID NO:4 having at least about 90% homology to a member selected from (SEQ ID NO:4, SEQ ED NO:4 positions 1- 200, SEQ ID NO:4 positions 1-400, SEQ ID NO: 4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5, SEQ ID NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725), and measuring an effect of the candidate compound modulator on the biological and/or pharmacological activity of the polypeptide. Preferred assays for modulators of nuclear transport receptors described herein fall into two general categories: 1) direct measurement of a biological activity as defined herein; and, 2) measurement of downstream events in the signaling cascade including cell/tissue/organism physiological manifestations. In another embodiment of the invention to identify agents which modulate a biological activity of the biomolecules recited herein, a nucleic acid sequence which encodes a transcription factor-selective nuclear transport receptor polypeptide as depicted may be ligated to a heterologous sequence to encode a fusion protein for use in a yeast 2- hybrid system. To screen compounds for the modulation of biological activity, it is necessary to encode a chimeric molecule as described herein for expression in hererologous host cells. Chimeric constructs are also used to express a 'bait', according to methods well known using a yeast two-hybrid system, using accessory native peptides that are expected to be associated with a nuclear transport receptor, e.g., SEQ ID NO:4. The two-hybrid system uses the ability of a pair of interacting proteins to bring a transcription activation domain into close proximity with a DNA-binding site that regulates the expression of an adjacent reporter gene. Compounds which are able to modulate the biological activity of a biomolecule as defined herein are identified by the their ability to effect protein:protein interactions (reconstitution of the chimeric transcriptional activators) and hence the yeast 2-hybrid readout assays well-known to artisans of ordinary skill in this area of molecular biology. Fields, S., et al, Trends Genet., 10:286 (1994); Allen, J.B., et al, TIBS, 20:511 (1995). Fields, S., Song, O., Nature 340:245 (1989). Commercially available systems such as the CLONTECH, Matchmaker™ systems and protocols may be used with the present invention. CLONTECH, Palo Alto, CA. See also, Mendelsohn, A.R., Brent, R., Curr. Op. Biotech., 5:482 (1994); Phizicky. E.M., Fields, S., Microbiological Rev., 59(1):94 (1995); Yang, M., et al., Nucleic Acids Res., 23(7): 1152 (1995); Fields, S., Sternglanz, R., TIG, 10(8):286 (1994); and US Patents 5,283,173, System to Detect Protein-Protein Interactions, and 5,468,614, which are incorporated herein by reference. Compounds which are identified generally according to methods described, contemplated, and referenced herein that modulate a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor polypeptide, e.g., the sequence as depicted in SEQ ED NO: 4 and/or SEQ ID NO: 5, are especially preferred embodiments of the present invention. An especially preferred embodiment of the present invention is a method for treatment of a patient in need of such treatment for a dysfunctional- transcription factor- selective nuclear transport receptor related condition which is mediated by a human transcription factor-selective nuclear transport receptor, e.g., SEQ ID NO:4, comprising administration of a therapeutically effective amount of a modulating compound which was identified by means of a method of identifying compounds that modulate a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor, comprising combining a candidate compound modulator with an isolated and purified polynucleotide molecule comprising a nucleic acid sequence which encodes a polypeptide comprising an amino acid sequence selected from (SEQ ID NO:3, SEQ ID NO:4, SEQ ED NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ED NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5, SEQ ID NO.5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ED NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725), and measuring an effect of the candidate compound modulator on the biological and/or pharmacological activity of the corresponding polypeptide. A method of modulating a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor polypeptide in a cell is preferred which comprises administering an effective amount of a polynucleotide comprising a sequence which encodes a polypeptide comprising a sequence as depicted by (SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5, SEQ ID NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725), to said cell. A method of modulating a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor in a cell, tissue, or organism is preferred which comprises administering an effective amount of a polynucleotide comprising a nucleic acid sequence derived from a nucleic acid which encodes SEQ ED NO:3, SEQ ID NO:4, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5, SEQ ED NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, or SEQ ID NO:5 positions 575-725 including but not limited to biologically-effective antisense molecules derived therefrom as well as dominant negative mutant versions. Compositions Pharmaceutically useful therapeutic compositions which comprise a nucleic acid contemplated herein, or a compound identified by means encompassed by the claims appended hereto that modulates the biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor, e.g., SEQ ED NO:4, may be formulated according to known methods such as by the admixture of a pharmaceutically acceptable carrier. Examples of such carriers and methods of formulation may be found in Remington's Pharmaceutical Sciences (Maack Publishing Co, Easton, PA). To form a pharmaceutically acceptable composition suitable for effective administration, such compositions will contain an effective amount of a polypeptide, nucleic acid, or compound modulator. Therapeutic compositions of the invention are administered to an individual or used in amounts sufficient to treat or diagnose transcription factor-selective nuclear transport receptor-related disorders. The effective amount may vary according to a variety of factors such as the individual's condition, weight, sex and age. Other factors include the mode of administration. Pharmaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose. The determination of an effective dose is well within the capability of those skilled in the art. The therapeutically effective dose can be estimated initially either in cell culture assays, eg, of neoplastic cells, or in animal models, usually mice, rabbits, dogs, or pigs. The animal model is also used to achieve a desirable concentration range and route of administration. Such information can then be used to determine useful doses and routes for administration in humans. A therapeutically effective dose refers to that amount of compound, peptide, or nucleic acid which ameliorate or prevent a dysfunctional apoptotic condition. The exact dosage is chosen by the individual physician in view of the patient to be treated. Compounds identified according to the methods disclosed herein as well as therapeutic nucleic acids and peptides contemplated herein may be used alone at appropriate dosages defined by routine testing in order to obtain optimal modulation of transcription factor-selective nuclear transport receptor activity. In addition, co- administration or sequential administration of these and other agents may be desirable. The pharmaceutical compositions may be provided to the individual by a variety of routes such as subcutaneous, topical, oral and intramuscular. Administration of pharmaceutical compositions is accomplished orally or parenterally. Methods of parenteral delivery include topical, intra-arterial (directly to the tissue), intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intraperitoneal, or intranasal administration. The present invention also has the objective of providing suitable topical, oral, systemic and parenteral pharmaceutical formulations for use in the novel methods of treatment of the present invention. The compositions containing compounds identified according to this invention as the active ingredient for use in the modulation of a transcription factor-selective nuclear transport receptor can be administered in a wide variety of therapeutic dosage forms in conventional vehicles for administration. For example, the compounds can be administered in such oral dosage forms as tablets, capsules (each including timed release and sustained release formulations), pills, powders, granules, elixirs, tinctures, solutions, suspensions, syrups and emulsions, or by injection. Likewise, they may also be administered in intravenous (both bolus and infusion), intraperitoneal, subcutaneous, topical with or without occlusion, or intramuscular form, all using forms well known to those of ordinary skill in the pharmaceutical arts. An effective but non-toxic amount of the compound, nucleic acid, or peptide desired can be employed as an the transcription factor-selective nuclear transport receptor modulating agent. The daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult human/per day. For oral administration, the compositions are preferably provided in the form of scored or unscored tablets containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, and 50.0 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. An effective amount of the drug is ordinarily supplied at a dosage level of from about 0.0001 mg/kg to about 100 mg/kg of body weight per day. The range is more particularly from about 0.001 mg/kg to 10 mg/kg of body weight per day. Even more particularly, the range varies from about 0.05 to about 1 mg/kg. Of course the dosage level will vary depending upon the potency of the particular compound. Certain compounds will be more potent than others. In addition, the dosage level will vary depending upon the bioavailability of the compound. The more bioavailable and potent the compound, the less compound will need to be administered through any delivery route, including but not limited to oral delivery. The dosages of transcription factor-selective nuclear transport receptor modulators are adjusted when combined to achieve desired effects. On the other hand, dosages of these various agents may be independently optimized and combined to achieve a synergistic result wherein the pathology is reduced more than it would be if either agent were used alone. Those skilled in the art will employ different formulations for nucleotides than for proteins or their inhibitors. Similarly, delivery of polynucleotides or polypeptides will be specific to particular cells and conditions. Examples Example I: Antibodies To prepare the A^bo antiserum, a DNA fragment from the mbo cDNA (SEQ ED NO: 1) (generated using Hindi and Kpnl) encoding SEQ ID NO:3 amino acids 505 to 702 was inserted in frame to the 6xHis coding sequence in pRSET C (cut with PvuLI and Kpnl; Invitrogen). The histidine-tagged Mbo fusion-protein was expressed inE. coli, purified on Ni-NTA resin (QIAGEN), and used for immunisation of rabbits at Agrisera (Umea, Sweden). The antiserum was affinity purified on Ni-NTA resin coupled with the histidine-tagged Mbo fusion protein and used at 1 : 100 dilution for Western blots and immunohistochemistry. Fixation was reduced to 15 minutes and the washes were shortened to 2x5 minutes followed by 2x30 minutes. Example II: Analysis of Immune Response To prepare larvae with low background levels of dorsal translocation, Drosophila eggs were collected on apple plates, dechorionated and transferred to new apple plates where they developed into larvae. The larvae fed on reconstituted dry yeast and were regularly transferred to new plates. For each experiment, larvae from the same plate were used. Injury was performed on third instar larvae by pricking the posterior part with a tungsten needle dipped in a mixed concentrated culture of E. coli and M. luteus. The infected larvae were analysed 45 min to 1 hour later for dorsal translocation, 2 hours later for antimicrobial peptide expression and 30 minutes later for cactus degradation. Dorsal translocation was analysed by antibody staining as described. Cactus degradation was assayed on western blots; whole larvae were turned inside-out, the gut was removed and the larval tissue transferred to sample buffer. After freezing and boiling, the samples were subjected to SDS-PAGE. Electrophoresis and immunoblotting were performed according to Sambrook et al. (1989). The polyclonal cactus antiserum (from S. Wassermann; Reach et al 1996, dev. biol 180) was used at 1/1000 dilution. Hybridising bands were detected with the Amersham ECL reagent according to the manufacturer. Antibacterial peptide induction was assayed by cecropin-LacZ expression using anti-beta galactosidase antibodies and by Northern blots of total RNA. Example HI: High Throughput Screening - Scintillation Proximity Assay (SPAi A high throughput screening assay is set up using Mbo, e.g., SEQ ID NO:3, expressed in either insect cells using the baculovirus expression system, or other mammalian cell lines such as CHO, Cos, or MEL. See, e.g., Needham M, et al, Protein Expr. Purif, 7(2): 173 (1996); Life Technologies, Gaithersburg, MD. Bac-to-Bac™ HT Baculovirus Expression System. Mbo can be tagged via expression as a fusion protein from commercially available expression vectors containing sequences, for example, FLAG, or c-myc/6-His to aid in purification and detection as is well-known in the art. Materials, purification and detection methods for FLAG-tag are available, for example, from Kodak, New Haven CT. Vectors, for use in the assay, include pFLAG.MAC for N-terminal expression of FLAG or pFLAG.CTC for C-terminal expression of FLAG (Kodak, New Haven CT). Purification method uses anti-FLAG M2 Affinity Gel. Detection is performed using monoclonal antibody Anti-FLAG M2 Monoclonal Antibody (# IB 13010) (Kodak, New Haven CT). C-myc/6-His-tag vectors, methods and detection reagents are obtained from Invitrogen Carlsbad CA: vector pcDNA3.1(-)/Myc-His Xpress™; and, purification: Xpress™ Purification System; and, detection: Anti-myc Antibody (# R950- 25) or Anti-His(C-term) Antibody (# R930-25). After expression and purification of the tagged Mbo the purified protein is fluorescently labeled through conjugation to BODIPY FL (Molecular Probes Eugene, OR) per the protocol provided by the manufacturer. Membranes of cells expressing Mbo are dissociated to release the protein by resuspending the membranes in 0.1 - 1% Triton X-100. Excess lipids are pelleted via centrifugation. Luo H, et al., Biochim Biophys Acta, 1052: 119 (1990); Reyl-Desmars F., et al., J.B.C., 264: 18789 (1989). Solubilized protein is bound to polylysine coated SPA beads (Amersham, Arlington Heights, IL) per the manufacturer's protocol. BODIPY FL-labeled Mbo is incubated alone and separately in the presence of candidate modulators with the protein-bound SPA beads. The beads are washed free of unbound BODIPY-Mbo. The relative amount of bound BODIPY-Mbo in each candidate sample is determined by measuring fluorescence emission at 513 nm (excitation wavelength = 505 nm) and compared to control sample emission. * * * All publications and patents mentioned in the above specification are herein incorporated by reference. Various modifications and variations of the described methods and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in molecular biology or related fields are intended to be within the scope of the following claims.

Claims: Claims 1. A purified polynucleotide molecule comprising a nucleic acid sequence which encodes a polypeptide having at least about 90% homology to a member selected from (SEQ DD NO:3, SEQ ED NO: 4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575- 725, SEQ ID NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ED NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575- 725). 2. A polynucleotide according to Claim 1 comprising a nucleic acid sequence which encodes a polypeptide selected from (SEQ ED NO:3, SEQ ED NO:4 positions 1-200, SEQ ED NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ ID NO:5 positions 550- 740, and SEQ ED NO:5 positions 575-725). 3. A polynucleotide of Claim 2 wherein the polynucleotide sequence comprises the sequence as depicted in SEQ ID NO: 1 or SEQ ID NO:2. 4. An expression vector comprising the polynucleotide of Claim 1. 5. A host cell transformed with the expression vector of Claim 4. 6. A purified polypeptide comprising the amino acid sequence as depicted in SEQ ID NO: 3 or a variant of SEQ ID NO: 3 having at least about 90% homology to a member selected from (SEQ ID NO:3, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1- 400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725). 7. A method for producing a polypeptide according to Claim 6, said method comprising the steps of: a) culturing a host cell according to Claim 5 under conditions suitable for the expression of said polypeptide. 8. A method of identifying compounds that modulate a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor polypeptide, comprising: (a) combining a candidate compound modulator with a polypeptide comprising a sequence having at least about 90% homology to a member selected from (SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO: 4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ID NO:4 positions 575-725, SEQ ID NO:5, SEQ ID NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ED NO:5 positions 50-350, SEQ ID NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725), and (b) measuring an effect of the candidate compound modulator on the biological and/or pharmacological activity of the polypeptide. 9. A method of identifying compounds that modulate a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor polypeptide according to Claim 8, comprising: (a) combining a candidate compound modulator with a host-cell which expresses said polypeptide, and (b) measuring an effect of the candidate compound modulator on the biological and/or pharmacological activity of the polypeptide. 10. A compound that modulates the activity of a transcription factor-selective nuclear transport receptor polypeptide identified by the method of Claim 9. 11. A method of modulating a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor polypeptide in a cell comprising administering an effective amount of a compound according to Claim 10 to said cell. 12. A method of modulating a biological and/or pharmacological activity of a transcription factor-selective nuclear transport receptor polypeptide in a cell comprising administering an effective amount of a polynucleotide comprising a sequence which encodes a polypeptide comprising a sequence as depicted by (SEQ ID NO:3, SEQ ED NO:4, SEQ ID NO:4 positions 1-200, SEQ ID NO:4 positions 1-400, SEQ ID NO:4 positions 50-350, SEQ ID NO:4 positions 550-740, SEQ ED NO:4 positions 575-725, SEQ ID NO:5, SEQ ED NO:5 positions 1-200, SEQ ID NO:5 positions 1-400, SEQ ID NO:5 positions 50-350, SEQ DD NO:5 positions 550-740, and SEQ ID NO:5 positions 575-725), to said cell.

Inventors: SAMAKOVLIS CHRISTOS, UV ANNE ELISABETH, SAMAKOVLIS, CHRISTOS, UV, ANNE, ELISABETH
Applicants: INNATE PHARMACEUTICALS AB, SAMAKOVLIS CHRISTOS, UV ANNE ELISABETH, SAMAKOVLIS, CHRISTOS, UV, ANNE, ELISABETH
Application year: 2000
Main classification: C07K14
IPC codes: A61K38, C07K14