HERPES SIMPLEX VIRUS ORIGIN BINDING PROTEIN N-TERMINAL TRUNCATION (OBP-NT)

Abstract: The invention provides herpes virus OBP-NT polypeptides and polynucleotides encoding OBP-NT polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are methods for utilizing OBP-NT polypeptides to screen for antiviral compounds.

Description: Herpes simplex virus Origin binding protein - N-terminal truncation (OBP-NT) FIELD OF THE INVENTION This invention relates to polynucleotides encoding a newly identified polypeptide, and their production and uses, as well as their vaπants, agonists and antagonists, and their uses In particular, the invention relates to polynucleotides encoding the polypeptide hereinafter referred to as herpes simplex virus oπgin binding protein N-terminal truncation, or "OBP-NT" BACKGROUND OF THE INVENTION The herpesviruses make up a medically important family of viruses comprised of three subfamilies, the alphaherpesviruses, betaherpesviruses and gammaherpesviruses These viruses are known to be able to produce infectious virus in a susceptible cell, as well as to remain latent in the host, reactivating in response to stimuli Herpes simplex virus types 1 and 2 (HSV-1 HSV-2) are alphaherpesviruses known to cause several types of disease in humans, including, for example, keratitis leading to blindness, encephahtisand herpes labiahs from HSV- 1 infection, neonatal disease and genital herpes from HSV-2 infection HSV-1 and HSV-2 are highly homologous viruses which were not distinguished until 1962 when the 2 serotypes were identified Since its isolation more than 70 years ago, HSV-1 has been the most intensively studied herpesvirus and is the prototype of the alphaherpesvirus subfamily Sequencing of the HSV-1 genome was completed in 1988, at which time 72 genes encoding 70 unique proteins were identified Since this time an additional 4 genes have been conclusively identified, with another 6 genes postulated HSV-2 has recently been sequenced, with identification of the analogous 74 unique proteins It is likely that additional as yet unknown proteins are encoded within the HSV-1 and, by analogy, HSV-2 genomes Despite the vast amount of research with HSV- 1 , many questions concerning the biology of this virus and of HSV-2 remain It is particularly preferred to employ HSV-1 and HSV-2 genes and gene products as targets for the development of antiviral reagents HSV-1 infects 40 to 80% of the general population, while HSV-2 infects 20 to 65% of the population HSV-2 infection is particularly significant in the prostitute and homosexual populations, where the frequency may approach 90% Current antivirals show only partial efficacy with reductions in pain, lesion seventy and shedding A medical need exists, therefore, for antivirals demonstrating improvement in overall efficacy, with decreased time to healing In addition, the ability to impact viral reactivation from latency would be a significant advantage and may have disease-modifying implications. Clearly, there exists a need for factors, such as the OBP-NT embodiment of the invention, that have a present benefit of being useful to screen compounds for antiviral activity Such factors are also useful to determine their role in pathogenesis of infection, dysfunction and disease. There is also a need for identification and characteπzation of such factors and their antagonists and agonists to find ways to prevent, ameliorate or correct such infection, dysfunction and disease. The origin binding protein (OBP), encoded within the UL9 gene of the HSV-1 genome, was originally identified in 1986 and further characterized in 1988, when binding activity to 2 homologous domains (site I and site II. also called Box I and Box II) within viral origins of DNA replication was described (Elias, P et al . 1986. Proc. Natl. Acad. Sci. USA 83:6322-6326, Elias, P. and I. R Lehman. 1988. Proc. Nat'l. Acad. Sci. USA 85.2959- 2963) Three viral origins of DNA replication are contained within the HSV-1 and HSV-2 genomes, 2 copies of oπS and 1 copy of oπL (Spaete. R R and Frenkel, N. 1985. Proc. Natl Acad. Sci. USA 82.694-698. Stow, N. D. and E C McMonagle. 1983 Virology 130:427-438, Dolan. A. et al. 1998 J Virol. 72.2010-2021 ) OπS contains 1 copy each of OBP binding sites I and II. as well as a third homologous sequence site III and an AT rich stretch; all of these domains are required for efficient origin function (see figure 1 , Elias, P. et al. 1990. J. Biol. Chem. 265 17167-17173). OπL contains 2 copies each of sites I and III and an AT rich stretch, which are required for efficient origin function (Lockshon, D and Galloway, D A 1988. Mol. Cell. Biol. 8.4018-4027) Despite considerable work in the field, the ability of the OBP to interact with site III remains controversial; while purified OBP does not appear to interact with oligonucleotides comprising site III, OBP from infected cell extracts has been identified as being able to bind site III oligonucleotides (Ohvo, P. D. et al. 1988. Proc. Nat'l. Acad. Sci. USA 85.5414-5418, Bruckner, R. C. et al. 1991. J. Biol. Chem. 266:2669-2674; Hazuda, D. J. et al. 1991. J. Biol. Chem. 266.24621- 24626; Dabrowski, C. E. and P. A. Schaffer. 1991. J. Virol. 65:3140-3150). Further, punfied OBP has been suggested to interact with site III within the context of the complete viral origin oπS (Elias, P. et al, 1990. J. Biol. Chem. 265.17167-17173; Koff, A et al. 1991. J. Virol. 65:3284-3292; Elias. P. et al. 1992. J. Biol. Chem. 267 17424-17429). The OBP has been shown to be essential for viral growth and DNA replication by the generation of viral mutants (Malik, A. K. et al. 1992. Virology 190:702-715). OBP has also been tested in cell-based transient replication assays, which demonstrated the requirement _ for helicase activity and specific dimeπzation sequences encoded within the N-terminus, and the ability to bind origin sequences encoded in the C-terminal half of the protein, for viral DNA replication (Hazuda. D. J. et al. 1992. J. Biol. Chem. 267: 14309-14315; Malik, A. K. and S. K Weller 1996. J. Virol. 70:7859-7866; Lee, S. S.-K. and I. R. Lehman. 1997. Proc. Natl. Acad. Sci. USA 94:2838-2842) Subsequent work resulted in the characterization of nucleotides important for origin binding activity, and demonstrated the relationship between the origin binding ability of OBP and origin-directed DNA replication (Challberg. M. D 1991. Seminars in Virology 2.247-256, Hernandez, T. R. et al. 1991. J. Virol. 65: 1649- 1652; Dabrowski, C. E. et al. 1994. Mol. Cell. Biol. 14:2545-2555). The UL9 (OBP) open reading frame is encoded within a 5.0-5.2 kilobase RNA in HSV-1 OBP comprises the protein component of the protein. DNA complex designated 'complex B ' in electrophoretic mobility shift assays (EMS A) following incubation of HSV- 1 infected cell extracts and radiolabeled HSV- 1 origin sequences (Baradaran. K et al. 1996. J Virol. 70-5673-5679) Novel RNAs 6.3-6.4 and 4 0-4 1 kilobases in length have been described, which are also transcribed from the UL9 region of the HSV-1 genome (Deb, S P. et al. 1993. Biochem and Biophys Res. Comm. 193 617-623, Baradaran, K. et al. 1994. J. Virol 68 4251-4261). The 4 0-4 1 kilobase RNA, designated UL8.5, was shown to encode a novel origin binding protein. OBPC, which is translated in-frame with the C-terminal (DNA binding) half of OBP (see figure 2) The function of this protein is as yet unknown, although a role in the switch from DNA replication to packaging of the viral genome into the capsid has been postulated OBPC comprises the protein component of the proteιn:DNA complex designated complex A' by EMS A following incubation of HSV-1 infected cell extracts and radiolabeled HSV- 1 origin sequences (Baradaran, K. et al. 1996. J. Virol. 70:5673-5679) Complex A has been shown to bind to origin sites I and II; the inability to bind to site III was shown to be due to a single nucleotide difference between sites I and III in the OBP binding domain (Dabrowski, C. E. and P. A. Schaffer. 1991. J. Virol. 65:3140-3150). The 6.3-6 4 kilobase RNA, designated UL9.5, has not yet been mapped but has been postulated to encode a protein which overlaps the N-terminus of the OBP (Baradaran, K. et al. 1994. J. Virol. 68:4251-4261). SUMMARY OF THE INVENTION It is an object of the invention to provide polypeptides that have been identified as OBP-NT polypeptides from herpes viruses, including from HSV-1 OBP-NT, HSV-2 OBP- NT, and VZV OBP-NT It is a further object of the invention to provide polynucleotides that encode OBP-NT polypeptides, particularly polynucleotides that encode the polypeptide herein designated HSV-1 OBP-NT, HSV-2 OBP-NT and VZV OBP-NT In a particularly preferred embodiment of the invention the polynucleotide compπses a region encoding HSV-1 OBP-NT polypeptide compnsing a sequence set out in Table 1 [SEQ ID NO 1] which includes a full length gene, or a vaπant thereof In another particularly preferred embodiment of the invention there is a OBP-NT protein from HSV-1 comprising the amino acid sequence of Table 1 [SEQ ID NO 2], or a variant thereof As a further aspect of the invention there are provided isolated nucleic acid molecules encoding OBP-NT, particularly HSV-1 OBP-NT, including mRNAs, cDNAs, genomic DNAs Further embodiments of the invention include biologically, diagnostically, prophylactically, clinically or therapeutically useful variants thereof, and compositions compnsing the same In accordance with another aspect of the invention, there is provided the use of a polynucleotide of the invention for therapeutic or prophylactic purposes, in particular genetic immunization Among the particularly preferred embodiments of the invention are naturally occurnng alle c vanants of OBP-NT and polypeptides encoded thereby In another aspect of the invention there are provided biologically, diagnostically, prophylactically, clinically or therapeutically useful vanants of polypeptides of HSV-1 OBP- NT and compositions compnsing the same Among the particularly preferred embodiments of the invention are vanants of HSV-1 OBP-NT polypeptide encoded by naturally occurnng alleles of the OBP-NT gene In a preferred embodiment of the invention there are provided methods for producing the aforementioned OBP-NT polypeptides In accordance with yet another aspect of the invention, there are provided inhibitors to polypeptides of the inventions, useful as antiviral agents, including, for example, antibodies or inhibitors of interactions between polynucleotides and polypeptides In accordance with certain preferred embodiments of the invention, there are provided products, compositions and methods for assessing OBP-NT expression, treating disease, assaying genetic vanation, and administenng a OBP-NT polypeptide or polynucleotide to an organism to raise an immunological response against a virus, especially HSV-1 In accordance with certain preferred embodiments of this and other aspects of the invention there are provided polynucleotides that hybridize to OBP-NT polynucleotide sequences, particularly under stnngent conditions In certain preferred embodiments of the invention there are provided antibodies against OBP-NT polypeptides In other embodiments of the invention there are provided methods for identifying compounds which bind to or otherwise interact with and inhibit or activate an activity of a polypeptide or polynucleotide of the invention compnsing contacting a polypeptide or polynucleotide of the invention with a compound to be screened under conditions to permit binding to or other interaction between the compound and the polypeptide or polynucleotide to assess the binding to or other interaction with the compound, such binding or interaction being associated with a second component capable of providing a detectable signal in response to the binding or interaction of the polypeptide or polynucleotide with the compound, and determining whether the compound binds to or otherwise interacts with and activates or inhibits an activity of the polypeptide or polynucleotide by detecting the presence or absence of a signal generated from the binding or interaction of the compound with the polypeptide or polynucleotide In accordance with yet another aspect of the invention, there are provided OBP-NT agonists and antagonists, preferably virostatic or virocidal agonists and antagonists In a further aspect of the invention there are provided compositions compnsing a OBP-NT polynucleotide or a OBP-NT polypeptide for administration to a cell or to a multicellular organism Vanous changes and modifications within the spint and scope of the disclosed invention will become readily apparent to those skilled in the art from reading the following descnptions and from reading the other parts of the present disclosure BRIEF DESCRIPTION OF THE FIGURES Figure 1 represents the nucleotide sequence of the core OBP binding domain defined within site I, and the homologous DNAs of sites II and III, for oπS and oπL Figure 2 represents characterized motifs of the protein encoded by the UL9 mRNA, as well as the predicted proteins encoded by the UL8 75 and UL8 5 mRNAs in relation to the UL9 encoded protein Figure 3 represents an EMS A of HSV- 1 infected cell extracts harvested 12 hours post-infection, incubated with radiolabelled site I. II, III or III(I) DNAs Figure 4 represents an EMS A of uninfected (Mock) cell extracts, or HSV- 1 infected cell extracts harvested at increasing time post-infection, incubated with a) radiolabelled site I DNA, b) radiolabelled site II DNA, and c) radiolabelled site III DNA Figure 5 represents a Western blot of uninfected (lane M) cell extracts, HSV-1 infected cell extracts harvested at 6 or 18 hours post-infection (lanes 6h, 18h), or extracts from cells infected with recombinant baculovirus expressing OBP (lane OBP), incubated with polyclonal antibody generated against the N-terminus of OBP-NT DESCRIPTION OF THE INVENTION The invention relates to OBP-NT polypeptides and polynucleotides as described in greater detail below In particular, the invention relates to polypeptides and polynucleotides of OBP-NT of HSV- 1 , which is related by amino acid sequence homology to other herpes virus OBP-NT proteins including, lor example, HSV-2 OBP-NT and VZV OBP-NT The invention relates especially to HSV-1 OBP-NT having the nucleotide and ammo acid sequences set out in Table 1 as SEQ ID NO 1 and SEQ ID NO 2 respectively TABLE 1 OBP-NT Polynucleotide and Polypeptide Sequences (A)Sequences from HSV-1 OBP-NT polynucleotide sequence [SEQ ID NO 1] 5-ATGAACGACCGCCCCTTCCACCGACTTATCGTCCAGGTGGAAAGCCTTCATCGCGTGGGC CCCAACCTTCTGAACAACTACGACGTCCTCGTTCTGGACGAGGTTATGTCGACGCTGGGC CAGCTCTATTCGCCAACGATGCAGCAACTGGGCCGCGTGGATGCGTTAATGCTACGCCTG CTGCGCATCTGTCCTCGGATCATCGCCATGGACGCAACCGCCAACGCGCAGTTGGTGGAC TTCCTGTGCGGTCTCCGGGGCGAAAAAAACGTGCATGTGGTGGTCGGCGAGTACGCCATG CCCGGGTTTTCGGCGCGCCGGTGCCTGTTTCTCCCGCGTCTGGGGACCGAGCTCCTGCAG GCTGCCCTGCGCCCGCCCGGGCCGCCGAGCGGCCCGTCTCCGGACGCCTCTCCGGAGGCC CGGGGGGCCACGTTCTTTGGGGAGCTGGAAGCGCGCCTTGGCGGGGGCGATAACATCTGC ATTTTTTCGTCGACGGTCTCCTTCGCGGAGATCGTGGCCCGGTTCTGCCGTCAGTTTACG GACCGCGTGCTGTTGCTTCACTCGCTCACCCCCCTCGGGGACGTGACCACGTGGGGCCAA TACCGCGTGGTTATATACACGACGGTCGTAACCGTGGGCCTCAGCTTCGATCCCCTGCAC TTTGATGGCATGTTCGCCTACGTGAAACCCATGAACTACGGACCGGACATGGTGTCCGTG TACCAGTCCCTGGGACGGGTGCGCACCCTCCGCAAGGGGGAGCTACTGATTTACATGGAC GGCTCCGGGGCGCGCTCGGAGCCCGTCTTTACGCCCATGCTCCTTAATCACGTGGTCAGT TCCTGCGGCCAGTGGCCCGCGCAGTTCTCCCAGGTCACAAACCTGCTGTGTCGCCGGTTC AAGGGGCGCTGTGACGCGTCGGCATGCGACACGTCGCTGGGGCGGGGGTCGCGCATCTAC AACAAATTCCGTTACAAACACTACTTTGAGAGATGCACGCTGGCGTGTCTCTCGGACAGC CTTAACATCCTTCACATGCTGCTGACCCTAAACTGCATACGCGTGCGCTTCTGGGGACAC GACGATACCCTGACCCCAAAGGACTTCTGTCTGTTTTTGCGGGGCGTACATTTCGACGCC CTCAGGGCCCAGCGCGATCTACGGGAGCTGCGGTGCCGGGATCCCGAGGCGTCGCTGCCG GCCCAGGCCGCCGAGACGGAGGAGGTGGGTCTTTTCGTCGAAAAATACCTCCGGTCCGAT GTCGCGCCGGCGGAAATTGTCGCGCTCATGCGCAACCTCAACAGCCTGATGGGACGCACG CGGTTTATTTACCTGGCGTTGCTGGAGGCCTGTCTCCGCGTTCCCATGGCCACCCGCAGC AGCGCCATATTTCGGCGGATCTATGACCACTACGCCACGGGCGTCATCCCCACGATCAAC GTCACCGGAGAGCTGGAGCTCGTGGCCCTGCCCCCCACCCTGAACGTAACCCCCGTCTGG GAGCTGTTGTGCCTGTGCAGCACCATGGCCGCGCGCCTGCATTGGGACTCGGCGGCCGGG GGATCTGGGAGGACCTTCGGCCCCGATGACGTGCTGGACCTACTGACCCCCCACTACGAC CGCTACATGCAGCTGGTGTTCGAACTGGGCCACTGTAACGTAACCGACGGACTTCTGCTC TCGGAGGAAGCCGTCAAGCGCGTCGCCGACGCCCTAAGCGGCTGTCCCCCGCGCGGGTCC GTTAGCGAGACGGACCACGCGGTGGCGCTGTTCAAGATAATCTGGGGCGAACTGTTTGGC GTGCAGATGGCCAAAAGCACGCAGACGTTTCCCGGGGCGGGGCGCGTTAAAAACCTCACC AAACAGACAATCGTGGGGTTGTTGGACGCCCACCACATCGACCACAGCGCCTGCCGGACC CACAGGCAGCTGTACGCCCTGCTTATGGCCCACAAGCGGGAGTTTGCGGGCGCGCGCTTC AAGCTACGCGTGCCCGCGTGGGGGCGCTGTTTGCGCACGCACTCATCCAGCGCCAACCCC AACGCTGACATCATCCTGGAGGCGGCGCTGTCGGAGCTCCCCACCGAGGCCTGGCCCATG ATGCAGGGGGCGGTGAACTTTAGCACCCTATAA-3 ' (B)Herpes simplex virus type 1 (HSV- 1 ) OBP-NT polypeptide sequence deduced from the polynucleotide sequence in this table [SEQ ID NO:2]. NH_-MNDRPFHRLIVQVESLHRVGPNLLNNYDVLV DEVMSTLGQLYSPTMQQLGRVDALMLRL LRICPRIIA DATANAQLVDFLCGLRGEK_NVHλ/VVGEYAMPGFΞARRC F PR GTELLQ AALRPPGPPSGPSPDASPEARGATFFGELEARLGGGDNICIFSSTVSFAEIVARFCRQFT DRVLLLHSLTP GDV TWGQYRWIYTTWTVGLSFDP HFDGMFAYVKPMNYGPD VSV YQSLGRVRTLRKGELLIYMDGSGARSEPVFTPMLLNHWSSCGQWPAQFSQVTN CRRF KGRCDASACDTSLGRGSRIYNKFRYKHYFERCTLACLSDSLNILHM LTLNCIRVRFWGH DDTLTPKDFCLFLRGVHFDALRAQRDLRELRCRDPEASLPAQAAETEEVGLFVEKYLRSD VAPAEIVALMR LNSLMGRTRFIYLALLEACLRVPMATRΞSAIFRRIYDHYATGVIPTIN VTGELELVALPPTLNVTPV ΞLLCLCSTMAAR H DSAAGGSGRTFGPDDVLDL TPHYD RYMQ VFE GHCNVTDGLL SEEAVKRVADALΞGCPPRGSVΞETDHAVALFKII GE FG VQMAKSTQTFPGAGRVKNLTKQTIVGLLDAHHIDHSACRTHRQLYA LMAHKREFAGARF KLRVPAWGRCLRTHSSSANPNADIILEAALSE PTEAWPMMQGAVNFSTL*-COOH (C)Polynucleotide sequences comprising Heφes simplex virus type 2 (HSV-2) OBP-NT ORF sequence [SEQ ID NO:3]. 5 ' - ATGAACGACCGCCCCTTCCACCGTCTCATCGTGCAGGTGGAAAGTCTTCATCGCGTGGGC CCGAACCTGTTGAACAACTACGACGTGCTCGTCACGAAGTCACTCATGTCGACGTTGGGC CAACTGTACTCGCCGACGATGCAGCAGCTGGGCCGCGTCGACGCGTTGATGCTGCGCCTG CTGCGCACGTGCCCGCGGATCATCGCCATGGACGCCACCGCCAACGCGCAGCTGGTGGAC TTTCTGTGCAGCCTCCGGGGCGAAAAGAACGTTCACGTGGTCATCGGGGAGTACGCCATG CCCGGATTTTCGGCGCGCCGTTGTCTGTTTCTCCCGCGCCTGGGGCCCGAGGTCCTGCAG GCGGCCCTGCGCCGCCGGGGGCCGGCGGGCGGGGCGCCCCCCCCGGACGCCCCCCCGGAC GCCACCTTCTTCGGGGAGCTGGAGGCGCGCCTGGCCGGCGGAGATAACGTCTGCATCTTC TCGTCGACGGTCTCCTTCGCGGAGGTCGTTGCCAGGTTCTGCCGGCAGTTTACGGACCGC GTGCTGCTGCTCCACTCGCTCACCCCGCCCGGCGACGTGACCACATGGGGCCGGTACCGG GTGGTCATCTACACAACGGTCGTGACGGTGGGCCTTAGCTTCGATCCGCCGCACTTTGAC AGCATGTTCGCCTACGTGAAACCCATGAACTACGGGCCGGACATGGTGTCCGTGTACCAG TCGCTGGGGCGGGTACGGACTCTCCGCAAGGGGGAGCTGCTGATCTACATGGACGGGTCC GGGGCGCGCTCGGAGCCCGTCTTTACGCCCATGCTGCTCAACCACGTGGTGAGCGCCAGC GGGCAGTGGCCGGCACAGTTTTCCCAGGTGACGAACCTGTTGTGCCGCCGGTTCAAAGGG CGCTGCGACGCGTCGCACGCCGACGCGGCGCAGGCGCGGGGGTCGCGCATCTACAGCAAA TTCCGGTACAAGCACTACTTCGAGAGGTGCACGCTGGCGTGCCTCGCGGACAGTCTTAAC ATCCTCCACATGCTTCTGACCCTCAACTGCATGCACGTGCGGTTCTGGGGCCACGACGCC GCGCTGACCCCGAGGAACTTTTGTCTGTTTTTGCGGGGGATACATTTTGATGCCCTGAGG GCCCAGCGGGATCTGCGGGAGCTGCGCTGCCAGGACCCCGACACGTCCCTGTCGGCCCAG GCCGCCGAGACGGAGGAGGTGGGCCTTTTCGTCGAAAAGTACCTCCGGCCGGACGTCGCG CCGGCCGAGGTGGTCGCGCTCATGCGCGGCCTCAACAGCCTGGTCGGCCGCACGCGGTTC ATCTACCTGGTGCTGCTGGAGGCCTGTCTTCGCGTCCCCATGGCCGCCCATAGCAGCGCC ATCTTCCGGCGGCTTTACGACCACTACGCCACGGGCGTCATCCCCACGATCAACGCCGCC GGAGAGCTGGAGCTTGTGGCCCTACACCCCACCCTAAACGTCGCCCCCGTCTGGGAGCTG TTCCGTCTGTGCAGCACCATGGCCGCGTGCCTGCAGTGGGACTCGATGGCCGGGGGGTCG GGGCGAACCTTTAGCCCCGAGGACGTGCTGGAGCTGCTGAACCCCCACTACGACCGCTAC ATGCAGCTGGTGTTCGAACTGGGCCACTGTAACGTGACCGACGGCCCCTTGCTGTCGGAG GACGCGGTTAAGCGCGTGGCCGACGCCCTGAGCGGCTGCCCCCCGCGCGGGTCCGTGAGC GAGACGGAGCACGCGCTGTCGCTGTTCAAGATCATCTGGGGCGAACTGTTCGGGGTGCAG CTGGCCAAGAGCACGCAGACGTTTCCCGGGGCGGGGCGCGTTAAAAACCTCACCAAGCGA GCCATCGTGGAGCTGCTGGACGCCCACCGCATCGACCACAGCGCCTGCCGGACGCACAGA CAGCTGTACGCGCTGCTGATGGCCCATAAGCGGGAGTTTGCGGGCGCGCGCTTCAAGCTG CGCGCGCCCGCGTGGGGGCGCTGCTTGCGCACGCACGCCTCCGGCGCCCAGCCCAACACT GACATCATTCTCGAGGCGGCTCTGTCGGAGCTTCCCACCGAGGCCTGGCCCATGATGCAG GGGGCGGTGAACTTTAGCACCCTATAA-3 ' (D)Heφes simplex virus type 2 (HSV-2) OBP-NT polypeptide sequence deduced from the polynucleotide ORF sequence [SEQ ID NO:4]. NH2-M DRPFHRLIVQVESLHRVGPNLL NYDVLVTKS MSTLGQLYSPT QQLGR'\DALMLRL LRTCPRIIAMDATANAQLVDFLCSLRGEK1WHVVIGEYAMPGFSARRC FLPRLGPEVLQ AALRRRGPAGGAPPPDAPPDATFFGELEARLAGGDNVCIFSSTVSFAEWARFCRQFTDR VL LHSLTPPGDVTTWGRYRWIYTTλ/VTVGLSFDPPHFDSMFAYVKPMNYGPDMVSVYQ SLGRVRTLRKGELLIYMDGSGARSEPVFTPMLLNHλΛSASGQWPAQFSQVTNLLCRRFKG RCDASHADAAQARGSRIYSKFRYKHYFERCTLACLADS NI HMLLTLNCMHVRFWGHDA A TPRNFC F RGIHFDA RAQRD RELRCQDPDTSLSAQAAETEEVG FVEKYLRPDVA PAEWA MRG NS VGRTRFIYLVLLEAC RVPMAAHSSAIFRRLYDHYATGVIPTINAA GE E VALHPTLNVAPV ELFRLCSTMAACLQV SMAGGSGRTFSPEDVLELLNPHYDRY MQ VFELGHCNVTDGPLLSEDAVKRVADALSGCPPRGSVSETEHALS FKIIWGE FGVQ LAKSTQTFPGAGRVKNLTKRAIVELLDAHRIDHSACRTHRQ YALLMAHKREFAGARFKL RAPAWGRCLRTHASGAQPNTDIILEAALSELPTEAWPMMQGAλ/NFST *-COOH (E) Heφes virus VZV OBP-NT polynucleotide sequence [SEQ ID NO:5]. 5 ' -ATGGGTTTTAAACGTTTGATTGTGCAACTTGAAAGCCTACACCGCGTATCCAGCGAAGCT ATCGACAGCTACGACGTATTAATACTGGATGAGGTAATGTCAGTGATTGGACAATTATAC TCCCCCACAATGAGACGTCTTTCCGCGGTTGATAGCCTATTATATCGTCTTTTAAATCGC TGTTCTCAAATTATCGCGATGGATGCTACAGTAAACTCGCAGTTTATTGATTTAATCTCC GGATTGCGTGGAGATGAAAACAT'ACACACAATTGTGTGTACATACGCGGGAGTTGGGTTC TCCGGAAGAACTTGCACGATCCTGCGTGATATGGGCATCGACACGCTTGTGCGAGTCATT AAACGATCTCCTGAACACGAGGATGTACGTACCATACACCAACTACGTGGAACATTTTTT GACGAACTAGCACTACGATTACAATGTGGGCATAACATCTGTATATTTTCATCAACTTTA TCGTTTTCGGAGCTAGTTGCTCAGTTTTGTGCAATATTTACAGACTCTATTCTTATTTTA AACTCAACTCGGCCCCTATGTAATGTAAACGAATGGAAACATTTTCGCGTGTTGGTGTAC ACTACCGTCGTGACCGTTGGATTGAGTTTTGACATGGCTCATTTTCATAGCATGTTTGCT TACATAAAGCCAATGTCATATGGGCCGGATATGGTATCGGTCTACCAGTCATTAGGGCGT GTACGTTTATTGCTACTTAATGAAGTTTTGATGTACGTCGATGGCTCAAGGACCAGATGC GGACCCCTGTTCTCGCCAATGTTACTAAACTTTACCATCGCAAATAAATTTCAATGGTTT CCTACACACACCCAAATAACTAACAAACTGTGCTGTGCATTTAGGCAACGATGTGCAAAT GCATTTACACGCTCGAACACCCATCTCTTCTCAAGATTTAAATACAAACACCTTTTCGAG AGATGCTCTCTTTGGAGTTTAGCCGATAGCATTAATATCTTACAAACTCTTTTGGCCTCT AACCAAATTTTGGTTGTATTGGATGGCATGGGTCCAATAACGGACGTTTCCCCAGTTCAA TTTTGTGCATTTATACACGATCTCAGACATAGCGCTAACGCCGTAGCTTCCTGTATGCGT TCTCTTAGACAGGACAATGACAGCTGCTTGACCGATTTTGGCCCTTCCGGATTTATGGCC GATAACATTACCGCGTTTATGGAAAAGTATCTTATGGAGTCAATTAATACCGAAGAACAA ATTAAAGTATTTAAAGCCCTTGCATGTCCAATAGAACAGCCTAGACTAGTCAATACGGCA ATATTGGGGGCGTGTATACGAATACCTGAAGCGTTGGAAGCATTTGACGTATTTCAAAAA ATATACACGCACTACGCTTCCGGTTGGTTTCCCGTCCTGGACAAAACCGGGGAATTTAGC ATCGCGACTATAACTACCGCCCCAAATTTAACCACACATTGGGAGCTGTTTCGCCGTTGT GCCTATATTGCAAAAACACTCAAGTGGAATCCGTCCACCGAAGGCTGTGTAACACAAGTT TTGGATACGGACATTAATACACTTTTCAATCAACACGGGGATTCGCTGGCTCAACTAATA TTTGAGGTTATGCGCTGTAACGTTACTGACGCTAAGATTATATTAAACCGCCCGGTTTGG CGAACAACCGGATTCTTAGATGGATGCCATAATCAATGCTTCCGTCCAATCCCTACAAAA CACGAATATAACATTGCTCTATTTCGTTTAATTTGGGAACAATTATTTGGCGCCCGCGTA ACTAAAAGTACCCAGACCTTTCCGGGAAGTACTCGTGTGAAAAACCTAAAAAAAAAAGAT CTAGAAACTTTACTTGATTCAATTAACGTGGATCGTTCTGCATGTCGTACCTACCGCCAG TTGTATAACCTGCTTATGAGCCAGCGCCATTCGTTCTCTCAACAGCGTTACAAAATTACT GCCCCCGCTTGGGCACGCCACGTGTATTTTCAAGCACATCAAATGCACTTGGCCCCGCAT GCCGAAGCCATGCTACAATTAGCGCTATCGGAACTGTCCCCGGGATCGTGGCCGCGGATA AACGGGGCGGTAAATTTTGAAAGTTTATAA-3 ' (F)VZV OBP-NT polypeptide sequence deduced from polynucleotide sequence [SEQ ID NO:6] NH2-MGFKR IVQLESLHRVSSEAIDSYDVLILDEVMSVIGQLYSPTMRRLSAVDSLLYR NR CSQIIAMDATVNSQFIDLISG RGDENIHTIVCTYAGVGFSGRTCTILRDMGIDTLVRVI KRSPEHEDVRTIHQLRGTFFDELALRLQCGHNICIFSSTLSFSELVAQFCAIFTDSILI NSTRPLCNVNE KHFRVLVYTTWTVGLSFDMAHFHSMFAYIKPMSYGPDMVSVYQSLGR W LLLNEVLMYVDGSRTRCGPLFSPMLLNFTIANKFQWFPTHTQITNKLCCAFRQRCAN AFTRSNTH FSRFKYKHLFERCS WSLADSINILQTLLASNQI W DGMGPITDVSPVQ FCAFIHDLRHSANAVASCMRS RQDNDSCLTDFGPSGFMADNITAFMEKY MESINTEEQ IKVFKALACPIEQPR VNTAILGACIRIPEALEAFDVFQKIYTHYASG FPVLDKTGEFS IATITTAPNLTTH ELFRRCAYIAKT K NPSTEGCVTQVLDTDINTLFNQHGDSLAQ I FEVMRCNVTDAKIILNRPVWRTTGFLDGCHNQCFRPIPTKHEYNIALFRLIWEQLFGARV TKSTQTFPGSTRVK LKKKDLETLLDSINVDRSACRTYRQLY LLMSQRHSFSQQRYKIT APA ARHVYFQAHQMHLAPHAEAMLQLA SELSPGSWPRINGAVNFESL*-COOH Polypeptides The polypeptides of the invention include polypeptides of Table 1 [SEQ ID NOS. 2, 4 and 6] (in particular the mature polypeptides) as well as polypeptides and fragments, particularly those which have the biological activity of OBP-NT, and also those which have at least 70% identity to a polypeptide of Table 1 or a relevant portion, preferably at least 80% identity to a polypeptide of Table 1 and more preferably at least 90% identity to a polypeptide of Table 1 and still more preferably at least 95% identity to a polypeptide of Table 1 and also including portions of such polypeptides with such portion of the polypeptide generally containing at least 30 amino acids and more preferably at least 50 amino acids. The invention also includes polypeptides of the formula X-(R 1 ) m -(R 2 )-(R 3 ) n -Y wherein, at the amino terminus, X is hydrogen or a metal, and at the carboxyl terminus, Y is hydrogen or a metal, Rj and R3 are any amino acid residue, m is an integer between 1 and 1000 or zero, n is an integer between 1 and 1000 or zero, and R ι^> an amino acid sequence of the invention, particularly an amino acid sequence selected from Table 1 In the formula above R 2 is onented so that its amino terminal residue is at the left, bound to R j and its carboxy terminal residue is at the nght, bound to R3 Any stretch ot ammo acid residues denoted by either R group, where m and/or n is greater than 1 , may be either a heteropolymer or a homopolymer, preferably a heteropolymer. A fragment is a vanant polypeptide having an amino acid sequence that entirely is the same as part but not all of the amino acid sequence of the aforementioned polypeptides As with OBP-NT polypeptides fragments may be "free-standing,' or comprised within a larger polypeptide of which they form a part or region, most preferably as a single continuous region, a single larger polypeptide Preferred fragments include, for example, truncation polypeptides having a portion of an amino acid sequence of Table 1, or of vanants thereof, such as a continuous senes of residues that includes the amino terminus, or a continuous senes of residues that includes the carboxyl terminus Degradation forms of the polypeptides of the invention in a host, particularly HSV-1, are also preferred Further preferred are fragments charactenzed by structural or functional attnbutes such as fragments that compnse alpha-helix and alpha-helix forming regions, beta-sheet and beta-sheet-forming regions, turn and tum-forming regions, coil and coil-forming regions, hydrophilic regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, flexible regions, surface-torming regions, substrate binding region, and high antigenic index regions Also preferred are biologically active fragments which are those fragments that mediate activities of OBP-NT, including those with a similar activity or an improved activity, or with a decreased undesirable activity. Also included are those fragments that are antigenic or lmmunogenic in an animal, especially in a human Particularly preferred are fragments compnsing receptors or domains of enzymes that confer a function essential for viability of HSV-1 or the ability to initiate, or maintain cause disease in an individual, particularly a human Vanants that are fragments of the polypeptides of the invention may be employed for producing the corresponding full-length polypeptide by peptide synthesis, therefore, these vanants may be employed as intermediates for producing the full-length polypeptides of the invention. In addition to the standard single and tnple letter representations for amino acids, the term "X' or "Xaa" may also be used in describing certain polypeptides of the invention "X" and "Xaa" mean that any of the twenty naturally occurring amino acids may appear at such a designated position in the polypeptide sequence Polynucleotides Another aspect of the invention relates to isolated polynucleotides, including the full length gene, that encode the OBP-NT polypeptide having a deduced amino acid sequence of Table 1 and polynucleotides closely related thereto and variants thereof Using the information provided herein, such as a polynucleotide sequence set out in Table 1 [SEQ ID NOS 1 , 3 and 5], a polynucleotide of the invention encoding OBP-NT polypeptide may be obtained using standard cloning and screening methods, such as those for cloning and sequencing chromosomal DNA fragments from virus using HSV- 1 KOS as starting matenal, followed by obtaining a full length clone For example, to obtain a polynucleotide sequence of the invention, such as a sequence given in Table 1 , a library of clones ot chromosomal DNA of HSV- 1 KOS in E.coli or some other suitable host may be probed with a radiolabeled ohgonucleotide, preferably a 17-mer or longer, denved from a partial sequence. Clones carrying DNA identical to that of the probe can then be distinguished using stnngent conditions Alternatively, a novel messenger RNA (mRNA) encoding a polypeptide can be identified by Northern blot analysis, probed with a radiolabeled ohgonucleotide or complementary RNA fragment, preferably a 17-mer or longer, derived from a partial sequence The 5 ' and 3' ends of the novel gene can then be mapped and the gene cloned By sequencing the individual clones thus identified with sequencing primers designed from the original sequence it is then possible to extend the sequence in both directions to determine the full gene sequence Conveniently, such sequencing is performed using denatured double stranded DNA prepared from a plasmid clone Suitable techniques are described by Mamatis, T . Fntsch, E F. and Sambrook et al., MOLECULAR CLONING, A LABORATORY MANUAL, 2nd Ed., Cold Spnng Harbor Laboratory Press, Cold Spnng Harbor, New York (1989) (see in particular Screening By Hybridization 1 90 and Sequencing Denatured Double-Stranded DNA Templates 13.70) Illustrative of the invention, a polynucleotide set out in Table 1 [SEQ ID NO: l] was discovered following Northern blotting of mRNA harvested following infection with HSV-1 The DNA sequences set out in Table 1 contain open reading frames encoding a protein having about the number of ammo acid residues set forth in Table 1 [SEQ ID NO:2] with a deduced molecular weight that can be calculated using amino acid residue molecular weight values well known the art The polynucleotide of SEQ ID NO 1 , between nucleotide number 1 and the stop codon which begins at nucleotide number 2131 of SEQ ID NO 1 , encodes the polypeptide of SEQ ID NO.2. OBP-NT of the invention is structurally related to other proteins of the heφesvirus family, particularly for example UL9 and UL8.5 of HSV-1 The invention provides a polynucleotide sequence identical over its entire length to a nucleotide coding sequence in Table 1 Also provided by the invention is the coding sequence for the mature polypeptide or a fragment thereof, by itself as well as the coding sequence for the mature polypeptide or a fragment in reading frame with other coding sequence, such as those encoding a leader or secretory sequence, a pre-, or pro- or prepro- protein sequence The polynucleotide may also contain non-cod g sequences, including for example, but not limited to non-coding 5 ' and 3' sequences, such as the transcnbed, non-translated sequences, termination signals, sequences that stabilize mRNA, introns, polyadenylation signals, and additional coding sequence which encode additional amino acids For example, a marker sequence that facilitates purification of the fused polypeptide can be encoded In certain embodiments of the invention, the marker sequence is a hexa-histidine peptide, as provided m the pQE vector (Qiagen, Inc.) and descnbed in Gentz et al, Proc Natl Acad. Sci., USA 86- 821-824 (1989), or an HA tag (Wilson et al, Cell 37: 767 (1984)) Polynucleotides of the invention also include, but are not limited to, polynucleotides compnsing a structural gene and its naturally associated sequences that control gene expression A preferred embodiment of the invention is a polynucleotide compnsing nucleotide 1 to the nucleotide immediately upstream of or including nucleotide 2131 set forth in SEQ ID NO l of Table 1, which encodes the HSV-1 OBP-NT polypeptide The invention also includes polvnucleotides of the formula X-(R 1 ) m -(R 2 HR3) π -Y wherein, at the 5 ' end of the molecule, X is hydrogen or a metal or together with Y defines a covalent bond, and at the 3' end of the molecule, Y is hydrogen or a metal or together with X defines the covalent bond, each occurrence of R j and R3 is independently any nucleic acid residue, m is an integer between 1 and 3000 or zero , n is an integer between 1 and 3000 or zero, and R 2 is a nucleic acid sequence of the invention, particularly a nucleic acid sequence selected from Table 1 In the polynucleotide formula above R is oriented so that its 5' end residue is at the left, bound to R j and its 3' end residue is at the right, bound to R3 Any stretch of nucleic acid residues denoted by either R group, where m and/or n is greater than 1, may be either a heteropolymer or a homopolymer, preferably a heteropolymer Where, in a preferred embodiment. X and Y together define a covalent bond, the polynucleotide of the above formula is a closed, circular polynucleotide. which can be a double-stranded polynucleotide wherein the formula shows a first strand to which the second strand is complementary In another preferred embodiment m and/or n is an integer between 1 and 1000. It is most preferred that the polynucleotides of the inventions are denved from HSV-1, however, they may preferably be obtained from organisms of the same taxonomic family The term polynucleotide encoding a polypeptide ' as used herein encompasses polynucleotides that include a sequence encoding a polypeptide of the invention, particularly a viral polypeptide and more a polypeptide of OBP-NT having an amino acid sequence set out in Table 1 The term also encompasses polynucleotides that include a single continuous region or discontinuous regions encoding the polypeptide (for example, interrupted by an insertion sequence or editing) together with additional regions, that also may contain coding and/or non-coding sequences The invention further relates to vanants of the polynucleotides descnbed herein that encode for vanants of the polypeptide having a deduced ammo acid sequences of Table 1 Vanants that are fragments of the polynucleotides of the invention may be used to synthesize full-length polynucleotides of the invention Further particularly preferred embodiments are polynucleotides encoding OBP-NT vanants, that have the amino acid sequence of OBP-NT polypeptides of Table 1 in which several, a few, 5 to 10, 1 to 5, 1 to 3, 2, 1 or no ammo acid residues are substituted, deleted or added, in any combination Especially preferred among these are silent substitutions, additions and deletions, that do not alter the properties and activities of OBP-NT Further preferred embodiments of the invention are polynucleotides that are at least 70% identical over their entire length to a polynucleotide encoding OBP-NT polypeptide having an ammo acid sequence set out in Table 1 and polynucleotides that are complementary to such polynucleotides Alternatively, most highly preferred are polynucleotides that compnse a region that is at least 80% identical over its entire length to a polynucleotide encoding OBP-NT polypeptide and polynucleotides complementary thereto In this regard, polynucleotides at least 90% identical over their entire length to the same are particularly preferred, and among these particularly prefened polynucleotides, those with at least 95% are especially preferred Furthermore, those with at least 97% are highly preferred among those with at least 95%, and among these those with at least 98% and at least 99% are particularly highly preferred, with at least 99% being the more preferred Preferred embodiments are polynucleotides that encode polypeptides that retain substantially the same biological function or activity as the mature polypeptide encoded by a DNA of Table 1 The invention further relates to polynucleotides that hybndize to the herein above- descnbed sequences In this legard, the invention especially relates to polynucleotides that hybndize under stnngent conditions to the herein above-descnbed polynucleotides As herein used, the terms "stnngent conditions" and "stnngent hybndization conditions mean hybndization will occur only if there is at least 95% and preferably at least 97% identity between the sequences An example of stringent hybridization conditions is overnight incubation at 42°C in a solution comprising 50% formamide, 5x SSC (150mM NaCl, 15mM tπsodium citrate), 50 mM sodium phosphate (pH7 6), 5x Denhardt's solution, 10% dextran sulfate. and 20 micrograms/ml denatured, sheared salmon sperm DNA, followed by washing the hybridization support in 0 lx SSC at about 65°C Hybridization and wash conditions are well known and exemplified in Sambrook. et al , Molecular Cloning A Laboratory Manual, Second Edition. Cold Spnng Harboi, N Y , (1989), particularly Chapter 1 1 therein The invention also provides a polynucleotide consisting essentially of a polynucleotide sequence obtainable by screening an appropriate library containing the complete gene for a polynucleotide sequence set forth in Table 1 under stringent hybndization conditions with a probe having the sequence of said polynucleotide sequence set forth in Table 1 or a fragment thereof, and isolating said DNA sequence Fragments useful for obtaining such a polynucleotide include, for example, probes and pnmers descnbed elsewhere herein As discussed additionally herein regarding polynucleotide assays of the invention, for instance, polynucleotides of the invention as discussed above may be used as a hybridization probe for RNA, cDNA and genomic DNA to isolate full-length cDNAs and genomic clones encoding OBP-NT and to isolate cDNA and genomic clones of other genes that have a high sequence identity to the OBP-NT gene Such probes generally will compnse at least 15 bases Preferably, such probes will have at least 30 bases and may have at least 50 bases Particularly preferred probes will have at least 30 bases and will have 50 bases or less For example, the coding region of the OBP-NT gene may be isolated by screening using a DNA sequence provided in Table 1 to synthesize an ohgonucleotide probe A labeled ohgonucleotide having a sequence complementary to that of a gene of the invention is then used to screen a library of cDNA, genomic DNA or mRNA to determine which members of the library the probe hybndizes to The polynucleotides and polypeptides of the invention may be employed, for example, as research reagents and matenals for discovery of treatments of and diagnostics for disease, particularly human disease, as further discussed herein relating to polynucleotide assays Polynucleotides of the invention that are oligonucleotides derived from the sequences of Table 1 may be used in the processes herein as described, but preferably for PCR, to determine whether or not the polynucleotides identified herein in whole or in part are transcribed by virus in infected tissue It is recognized that such sequences will also have utility in diagnosis of the stage of infection the pathogen has attained The invention also provides polynucleotides that may encode a polypeptide that is the mature protein plus additional amino or carboxvl-terminal amino acids, or amino acids mtenor to the mature polypeptide (when the mature form has more than one polypeptide chain, for instance) Such sequences may play a role in processing of a protein from precursor to a mature form, may allow protein transport, may lengthen or shorten protein half-life or may facilitate manipulation of a protein for assay or production, among other things As generally is the case in vivo, the additional amino acids may be processed away from the mature protein by cellular enzymes A precursor protein, ha\ ing the mature form of the polypeptide fused to one or more prosequences may be an inactive form of the polypeptide When prosequences are removed such inactive precursors generally are activated Some or all of the prosequences may be removed before activation Generally, such precursors are called proprotems In addition to the standard A, G, C, TAJ representations for nucleic acid bases, the term "N" may also be used in describing certain polynucleotides of the invention "N" means that any of the four DNA or RNA bases may appear at such a designated position in the DNA or RNA sequence, except it is preferred that N is not a base that when taken in combination with adjacent nucleotide positions, when read in the correct reading frame, would have the effect of generating a premature termination codon in such reading frame In sum, a polynucleotide of the invention may encode a mature protein, a mature protein plus a leader sequence (which may be referred to as a pro φrotein), a precursor of a mature protein having one or more prosequences that are not the leader sequences of a preprotein, or a preproprotein, which is a precursor to a proprotein, having a leader sequence and one or more prosequences, which generally are removed dunng processing steps that produce active and mature forms of the polypeptide Vectors, host cells, expression The invention also relates to vectors that compnse a polynucleotide or polynucleotides of the invention, host cells that are genetically engineered with vectors of the invention and the production of polypeptides of the invention by recombinant techniques Cell-free translation systems can also be employed to produce such proteins using RNAs denved from the DNA constructs of the invention For recombinant production, host cells can be genetically engineered to incoφorate expression systems or portions thereof or polynucleotides of the invention Introduction of a polynucleotide into the host cell can be effected by methods descnbed in many standard laboratory manuals, such as Davis et al., BASIC METHODS IN MOLECULAR BIOLOGY, (1986) and Sambrook et al , MOLECULAR CLONING A LABORATORY MANUAL. 2nd Ed , Cold Spnng Harbor Laboratory Press, Cold Spnng Harbor. N Y ( 1989), such as, calcium phosphate transfection, DEAE-dextran mediated transfection, transvection, microinjection, cationic hpid-mediated transfection, electroporation, transduction, scrape loading, ballistic introduction and infection Representative examples of appropnate hosts include bactenal cells, such as streptococci, staphylococci, enterococci E coh, streptomyces and Bacillus subtihs cells, fungal cells, such as yeast cells and Aspergillus cells; insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS, HeLa, C127, 3T3, BHK, 293, Vero and Bowes melanoma cells, and plant cells. A great vanety of expression systems can be used to produce the polypeptides of the invention Such vectors include, among others, chromosomal, episomal and virus-denved- vectors, e g , vectors denved from bactenal plasmids, from bactenophage, from transposons, from yeast episomes, from insertion elements, from yeast chromosomal elements, from viruses such as heφesviruses, baculoviruses, papova viruses, such as SV40, vaccinia viruses, adenoviruses, fowl pox viruses, pseudorabies viruses and retroviruses, and vectors denved from combinations thereof, such as those denved from plasmid and bactenophage genetic elements, such as cosmids and phagemids. The expression system constructs may contain control regions that regulate as well as engender expression Generally, any system or vector suitable to maintain, propagate or express polynucleotides and/or to express a polypeptide m a host may be used for expression in this regard. The appropriate DNA sequence may be inserted into the expression system by any of a vanety of well-known and routine techniques, such as, for example, those set forth m Sambrook et al , MOLECULAR CLONING, A LABORATORY MANUAL, (supra). For secretion of the translated protein into the lumen of the endoplasmic reticulum, into the peπplasmic space or into the extracellular environment, appropriate secretion signals may be incoφorated into the expressed polypeptide. These signals may be endogenous to the polypeptide or they may be heterologous signals. Polypeptides of the invention can be recovered and purified from recombinant cell cultures by well-known methods including ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography, and lectin chromatography Most preferably, high performance liquid chromatography is employed for punfication Well known techniques for refolding protein may be employed to regenerate active conformation when the polypeptide is denatured dunng isolation and or punfication Diagnostic, Prognostic, Serotyping and Mutation Assays This invention is also related to the use of the OBP-NT polynucleotides of the invention for use as diagnostic reagents. Detection of OBP-NT in a eukaryote, particularly a mammal, and especially a human, will provide a diagnostic method for diagnosis of a disease Eukaryotes (herein also "ιndivιdual(s)"), particularly mammals, and especially humans, particularly those infected or suspected to be infected with an organism compnsing the HSV OBP-NT gene may be detected at the nucleic acid level by a vanety of techniques Nucleic acids for diagnosis may be obtained from an infected individual's cells and tissues, such as cells of neuronal ongin, blood, mucosa, epithelium, and skin. Genomic DNA may be used directly for detection or may be amplified enzymatically by using PCR or other. amplification technique pnor to analysis RNA, cDNA and genomic DNA may also be used m the same ways Using amplification, charactenzation of the species and strain of virus present in an individual, may be made by an analysis of the genotype of the viral gene Deletions and insertions can be detected by a change in size of the amplified product in companson to the genotype of a reference sequence Point mutations can be identified by hybndizing amplified DNA to labeled OBP-NT polynucleotide sequences Perfectly matched sequences can be distinguished from mismatched duplexes by RNase digestion or by differences in melting temperatures DNA sequence differences may also be detected by alterations in the electrophoretic mobility of the DNA fragments in gels, with or without denatunng agents, or by direct DNA sequencing See. e g , Myers et al , Science, 230 1242 (1985) Sequence changes at specific locations also may be revealed by nuclease protection assays, such as RNase and S 1 protection or a chemical cleavage method See, e g , Cotton et al . Proc Nad Acad Sci USA 85 4397-4401 (1985) Cells carrying mutations or polymoφhisms (allehc vanations) in the gene of the invention may also be detected at the DNA or RNA level by a vanety of techniques, to allow for serotyping, for example For example, RT-PCR can be used to detect mutations in the RNA It is particularly prefened to used RT-PCR in conjunction with automated detection systems, such as for example, GeneScan RNA, cDNA or genomic DNA may also be used for the same puφose PCR or RT-PCR As an example PCR pnmers complementary to a nucleic acid encoding OBP-NT can be used to identify and analyze mutations Examples of representative pnmers are shown below in Table 2 Table 2 Primers for amplification of OBP-NT polynucleotides SEQ ID NO PRIMER SEQUENCE 5 5'-CCTGTCGTCGGAGTTTTACCCAG-3' 6 5'-GAGGTGCGGTCCCGAGACTTATAG-3' The invention also includes pnmers of the formula X-(R 1 ) m -(R 2 )-(Rι) n -Y wherein, at the 5' end of the molecule, X is hydrogen or a metal, and at the 3' end of the molecule. Y is hydrogen or a metal, R ] and R3 is anv nucleic acid residue, m is an integer between 1 and 20 or zero , n is an integer between 1 and 20 or zero, and R is a pnmer- sequence of the invention, particularly a pnmer sequence selected from Table 2 In the polynucleotide formula above R is onented so that its 5' end residue is at the left, bound to R\ and its 3' end residue is at the right, bound to R Any stretch of nucleic acid residues denoted by either R group, where m and/or n is greater than 1 , may be either a heteropolymer or a homopolymer, preferably a heteropolymer being complementary to a region of a polynucleotide of Table 1 In a pretened embodiment m and/or n is an integer between 1 and 10. The invention further provides these pnmers with 1 , 2, 3 or 4 nucleotides removed from the 5' and/or the 3' end These pnmers may be used for, among other things, amplifying OBP-NT DNA isolated from a sample denved from an individual The pnmers may be used to amplify the gene isolated from an infected individual such that the gene may then be subject to vanous techniques for elucidation of the DNA sequence In this way. mutations in the DNA sequence may be detected and used to diagnose infection and to serotype and/or classify the infectious agent The invention further provides a process for diagnosing disease, preferably viral infections, more preferably infections by HSV- 1 , comprising determining from a sample denved from an individual an increased level of expression of polynucleotide having a sequence of Table 1 Increased or decreased expression of OBP-NT polynucleotide can be measured using any one of the methods well known in the art for the quantitation of polynucleotides, such as, for example, amplification, PCR. RT-PCR. RNase protection. Northern blotting and other hybridization methods In addition, a diagnostic assay in accordance with the invention for detecting over- expression of OBP-NT protein compared to normal control tissue samples may be used to detect the presence of an infection, for example Assay techniques that can be used to determine levels of a OBP-NT protein, in a sample denved from a host are well-known to those of skill m the art Such assay methods include radioimmunoassays, competitive-binding assays, Western Blot analysis and ELISA assays The differences in the RNA or genomic sequence between viruses of diffenng phenotypes can also be determined If a mutation or sequence is observed in some or all of the viruses of a certain phenotype, but not in any viruses lacking that phenotype, then the mutation or sequence is likely to be the causative agent of the phenotype In this way, chromosomal regions may be identified that confer viral pathogenicity, growth charactenstics, survival characteπstics and/or ecological niche charactenstics Differential Expression The polynucleotides and polynucleotides of the invention may be used as reagents for differential screening methods There are many differential screening and differential display methods known in the art in which the polynucleotides and polypeptides of the invention may be used For example, the differential display technique is described by Chuang et al , J Bacteriol 175 2026-2036 (1993) This method identifies those genes which are expressed in an organism by identifying mRNA present using randomly-pπmed RT-PCR By comparing pre-infection and post infection profiles, genes up and down regulated dunng infection can be identified and the RT-PCR product sequenced and matched to ORF 'unknowns' RT-PCR may also be used to analyze gene expression patterns For RT-PCR using the polynucleotides of the invention, messenger RNA is isolated from virally infected tissue, and the amount of each mRNA species assessed by reverse transcription of the RNA sample primed with random hexanucleotides followed by PCR with gene specific primer pairs The determination of the presence and amount of a particular mRNA species by quantification of the resultant PCR product provides information on the viral genes which are transcribed in the infected tissue Analysis of gene transcription can be carried out at different times of infection to gain a detailed knowledge of gene regulation in viral pathogenesis allowing for a clearer understanding of which gene products represent targets for screens for antivirals Because of the gene specific nature of the PCR primers employed it should be understood that the viral mRNA preparation need not be free of mammalian RNA Optimally the viral mRNA is prepared from infected tissue by mechanical disruption in the presence of TRIzol (GIBCO-BRL) for very short penods of time, subsequent processing according to the manufacturers of TRIzol reagent and DNAase treatment to remove contaminating DNA Preferably the process is optimised by finding those conditions which give a maximum amount of HSV-1 RNA as detected by probing Northerns with a suitably labelled sequence specific ohgonucleotide or RNA probe Each of these techniques may have advantages or disadvantages depending on the particular application The skilled artisan would choose the approach that is the most relevant with the particular end use in mind Antibodies The polypeptides of the invention or variants thereof, or cells expressing them can be used as an immunogen to produce antibodies lmmunospecific for such polypeptides "Antibodies as used herein includes monoclonal and polyclonal antibodies, chimenc, single chain, simianized antibodies and humanized antibodies, as well as Fab fragments including the products of an Fab immunolglobu n expression library Antibodies generated against the polypeptides of the invention can be obtained by administenng the polypeptides or epitope-beanng fragments, analogues or cells to an animal, preferably a nonhuman, using routine protocols For preparation of monoclonal antibodies, any technique known in the art that provides antibodies produced by continuous cell line cultures can be used Examples include vanous techniques, such as those in Kohler, G and Milstein, C , Nature 256 495-497 (1975) Kozbor et al . Immunology 4 72 (1983), Cole et al , pg 77-96 in MONOCLONAL ANTIBODIES AND CANCER THLRAPY Alan R Liss, Inc (1985) Techniques for the production of single chain antibodies (U S Patent No 4,946,778) can be adapted to produce single chain antibodies to polypeptides of this invention Also, transgemc mice, or other organisms such as other mammals, may be used to express humanized antibodies Alternatively phage display technology may be utilized to select antibody genes with binding activities towards the polypeptide either from repertoires of PCR amplified v- genes of lymphocytes from humans screened for possessing anti-OBP-NT or from naive libraries (McCafferty, J et al ( 1990). Nature 348 552-554, Marks J et al , (1992) Biotechnology 10 779-783) The affinity of these antibodies can also be improved by chain shuffling (Clackson. T et al , ( 1991 ) Nature 352, 624-628) If two antigen binding domains are present each domain may be directed against a different epitope - termed antibodies The above-descnbed antibodies may be employed to isolate or to identify clones expressing the polypeptides to punfy the polypeptides by affinity chromatography Thus, among others, antibodies against OBP-NT-polypeptide may be employed to treat infections, particularly viral infections Polypeptide variants include antigenically, epitopically or immunologically equivalent variants that form a particular aspect of this invention The term antigenically equivalent derivative' as used herein encompasses a polypeptide or its equivalent which will be specifically recognized by certain antibodies which, when raised to the protein or polypeptide according to the invention, interfere with the immediate physical interaction between pathogen and mammalian host The term "immunologically equivalent denvative" as used herein encompasses a peptide or its equivalent which when used in a suitable formulation to raise antibodies in a vertebrate, the antibodies act to interfere with the immediate physical interaction between pathogen and mammalian host The polypeptide, such as an antigenically or immunologically equivalent derivative or a fusion protein thereof is used as an antigen to immunize a mouse or other animal such as a rat or chicken The fusion protein may provide stability to the polypeptide The antigen may be associated, for example by conjugation, with an immunogenic earner protein for example bovine serum albumin (BSA) or keyhole limpet haemocyamn (KLH) Alternatively a multiple antigenic peptide comprising multiple copies of the protein or polypeptide, or an antigenically or immunologically equivalent polypeptide thereof may be sufficiently antigenic to improve immunogenicity so as to obviate the use of a earner Preferably, the antibody or variant thereof is modified to make it less immunogenic in the individual For example, if the individual is human the antibody may most preferably be "humanized", where the complimentaπty determining regιon(s) of the hybπdoma-deπved antibody has been transplanted into a human monoclonal antibody , for example as described in Jones, P et al ( 1986), Natui e 321 , 522-525 or Tempest et al , ( 1991 ) Biotechnology 9, 266-273 The use of a polynucleotide of the invention in genetic immunization will preferably employ a suitable delivery method such as direct injection of plasmid DNA into muscles (Wolff et al , Hum Mol Genet 1992, 1 363 Manthoφe et al , Hum Gene Ther 1963 4, 419), delivery of DNA complexed with specific protein carriers (Wu et al , J Biol Chem 1989 264, 16985), coprecipitation of DNA with calcium phosphate (Benvenisty & Reshef, PNAS USA, 1986 83,9551 ), encapsulation of DNA various forms of liposomes (Kaneda et al , Science 1989 243,375), particle bombardment (Tang et al , Nature 1992, 356 152, Eisenbraun et al , DNA Cell Biol 1993, 12 791) and in vivo infection using cloned retroviral vectors (Seeger et al . PNAS USA 1984 81,5849) Antagonists and agonists - assays and molecules Polypeptides of the invention may also be used to assess the binding of small molecule substrates and gands in, for example, cells, cell-free preparations chemical branes, and natural product mixtures These substrates and hgands may be natural substrates and hgands or may be structural or functional mimetics See, e g , Co gan et al . Current Protocols in Immunology 1(2) Chapter 5 ( 1991) The invention provides a method of screening compounds to identify those which enhance (agonist) or block (antagonist) the action of OBP-NT polypeptides or polynucleotides, particularly those compounds that are virostatic and or virocidal The method of screening may involve high-throughput techniques For example, to screen for agonists or antagonists, a synthetic reaction mix, a cellular compartment, such as a nucleus, cytoplasm or membrane, or a preparation of any thereof, compnsing OBP-NT polypeptide and a labeled substrate or ligand of such polypeptide is incubated in the absence or the presence of a candidate molecule that may be a OBP-NT agonist or antagonist The ability of the candidate molecule to agonize or antagonize the OBP-NT polypeptide is reflected in decreased binding of the labeled ligand or decreased production of product from such substrate Molecules that bind gratuitously, i e . without inducing the effects of OBP-NT polypeptide are most likely to be good antagonists Molecules that bind well and increase the rate of product production from substrate are agonists Detection of the rate or level of production of product from substrate may be enhanced by using a reporter system Reporter systems that may be useful in this regard include but are not limited to colonmetnc labeled substrate converted into product, a reporter gene that is responsive to changes in OBP-NT polynucleotide or polypeptide activity, and binding assays known in the art Another example of an assay for OBP-NT antagonists is a competitive assay that combines OBP-NT and a potential antagonist with OBP-NT-binding molecules, recombinant OBP-NT binding molecules, natural substrates or hgands, or substrate or ligand mimetics, under appropπate conditions for a competitive inhibition assay OBP-NT can be labeled, such as by radioactivity or a colonmetnc compound, such that the number of OBP-NT molecules bound to a binding molecule or converted to product can be determined accurately to assess the effectiveness of the potential antagonists Potential antagonists include small organic molecules, peptides, polypeptides and antibodies that bind to a polynucleotide or polypeptide of the invention and thereby inhibit or extinguish its activity. Potential antagonists also may be small organic molecules, a peptide, a polypeptide such as a closely related protein or antibody that binds the same sites on a binding molecule, such as a binding molecule, without inducing OBP-NT-induced activities, thereby preventing the action of OBP-NT by excluding OBP-NT from binding Potential antagonists include a small molecule that binds to and occupies the binding site of the polypeptide thereby preventing binding to viral or cellular binding molecules, such that normal biological activity is prevented. Examples of small molecules include but are not limited to small organic molecules, peptides or peptide- ke molecules Other potential antagonists include antisense molecules (see Okano, J Neurochem 56 560 (1991), OLIGODEOXYNUCLEOTIDES AS ANTISENSE INHIBITORS OF GENE EXPRESSION, CRC Press, Boca Raton, FL ( 1988). for a descnption of these molecules) Preferred potential antagonists include compounds related to and vanants of OBP-NT Each of the DNA sequences provided herein may be used in the discovery and development of antiviral compounds The encoded protein, upon expression, can be used as a target for the screening of antiviral drugs Additionally, the DNA sequences encoding the ammo terminal regions of the encoded protein or translation facilitating sequences of the respective mRNA can be used to construct antisense sequences to control the expression of the coding sequence of interest The antagonists and agonists of the invention may be employed, for instance, to inhibit and treat diseases Vaccines Another aspect of the invention relates to a method for inducing an immunological response in an individual, particularly a mammal which compnses inoculating the individual with OBP-NT, or a fragment or variant thereof adequate to produce antibody and/ or T cell immune response to protect said individual from infection, particularly viral infection and most particularly HSV-1 infection Also provided are methods whereby such immunological response slows viral replication Yet another aspect of the invention relates to a method of inducing immunological response in an individual which comprises delivering to such individual a nucleic acid vector to direct expression of OBP-NT, or a fragment or a variant thereof for expressing OBP-NT, or a fragment or a variant thereof in vivo in order to induce an immunological response, such as. to produce antibody and/ or T cell immune response, including, for example, cytokine-producing T cells or cytotoxic T cells, to protect said individual from disease, whether that disease is already established within the individual or not One way of administering the gene is by accelerating it into the desired cells as a coating on particles or otherwise Such nucleic acid vector may compnse DNA, RNA, a modified nucleic acid, or a DNA/RNA hybrid A further aspect of the invention relates to an immunological composition which, when introduced into an individual capable or having induced within it an immunological response, induces an immunological response in such individual to a OBP-NT or protein coded therefrom, wherein the composition compnses a recombinant OBP-NT or protein coded therefrom comprising DNA which codes for and expresses an antigen of said OBP- NT or protein coded therefrom The immunological response may be used therapeutically or prophylactically and may take the form of antibody immunity or cellular immunity such as that arising from CTL or CD4+ T cells A OBP-NT polypeptide or a fragment thereof may be fused with co-protein which may not by itself produce antibodies, but is capable of stabilizing the first protein and producing a fused protein which will have immunogenic and protective properties This fused recombinant protein, preferably further compnses an antigenic co-protein, such as Glutathione-S-transferase (GST) or beta-galactosidase, relatively large co-proteins which solubihze the protein and facilitate production and purification thereof Moreover, the co- protein may act as an adjuvant in the sense of providing a generalized stimulation of the immune system The co-protem may be attached to either the amino or carboxy terminus of the first protein Provided by this invention are compositions, particularly vaccine compositions, and methods comprising the polypeptides or polynucleotides of the invention and immunostimulatory DNA sequences, such as those described m Sato, Y et al Science 273 352 (1996) The polypeptide may be used as an antigen for vaccination of a host to produce specific antibodies which protect against viral infection, for example by blocking viral replication or viral reactivation from latency The invention also includes a vaccine formulation which compnses an immunogenic recombinant protein of the invention together with a suitable carrier Since the protein may be broken down in the stomach, it is preferably administered parenterally, including, for example, administration that is subcutaneous, intramuscular, intravenous, or lntradermal Formulations suitable for parenteral administration include aqueous and non- aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteπostats and solutes which render the formulation insotomc with the bodily fluid, preferably the blood, of the individual, and aqueous and non-aqueous sterile suspensions which may include suspending agents or thickening agents The formulations may be presented in unit-dose or multi-dose containers, for example, sealed ampules and vials and may be stored in a freeze-dπed condition requiring only the addition of the stenle liquid earner immediately prior to use The vaccine formulation may also include adjuvant systems for enhancing the lmmunogenicity of the formulation, such as oil-in water systems and other systems known in the art The dosage will depend on the specific activity of the vaccine and can be readily determined by routine experimentation While the invention has been described with reference to certain OBP-NT protein, it is to be understood that this covers fragments of the naturally occurring protein and similar proteins with additions, deletions or substitutions w hich do not substantially affect the immunogenic properties of the recombinant protein Compositions, kits and administration The invention also relates to compositions compnsing the polynucleotide or the polypeptides discussed above or their agonists or antagonists The polypeptides of the invention may be employed in combination with a non-sterile or stenle earner or earners for use with cells, tissues or organisms, such as a pharmaceutical earner suitable for administration to a subject Such compositions comprise, for instance, a media additive or a therapeutically effective amount of a polypeptide of the invention and a pharmaceutically acceptable earner or excipient Such earners may include, but are not limited to, saline, buffered saline, dextrose, water, glycerol, ethanol and combinations thereof The formulation should suit the mode of administration The invention further relates to diagnostic and pharmaceutical packs and kits compnsing one or more containers filled with one or more of the ingredients of the aforementioned compositions of the invention Polypeptides and other compounds of the invention may be employed alone or in conjunction with other compounds, such as therapeutic compounds The pharmaceutical compositions may be administered in any effective, convenient manner including, for instance, administration by topical, oral, anal, vaginal, intravenous, intrapentoneal, intramuscular, subcutaneous, intranasal or intradermal routes among others In therapy or as a prophylactic, the active agent may be administered to an individual as an injectable composition, for example as a sterile aqueous dispersion, preferably lsotomc Alternatively the composition may be formulated for topical application for example in the form of ointments, creams, lotions, eye ointments, eye drops, ear drops, mouthwash, impregnated dressings and sutures and aerosols, and may contain appropriate conventional additives, including, for example, preservatives, solvents to assist drug penetration, and emollients in ointments and creams Such topical formulations may also contain compatible conventional carriers, for example cream or ointment bases, and ethanol or oleyl alcohol for lotions Such carriers may constitute from about 1 % to about 98% by weight of the formulation, more usually they will constitute up to about 80% by weight of the formulation For administration to mammals, and particularly humans, it is expected that the daily dosage level of the active agent will be from 0 01 mg/kg to 10 mg/kg, typically around 1 mg/kg The physician in any event will determine the actual dosage which will be most suitable for an individual and will vary with the age, weight and response of the particular individual The above dosages are exemplary of the average case There can, of course, be individual instances where higher or lower dosage ranges are mented, and such are within the scope of this invention A vaccine composition is conveniently in injectable form Conventional adjuvants may be employed to enhance the immune response A suitable unit dose for vaccination is 0 5-5 microgram kg of antigen, and such dose is preferably administered 1-3 times and with an interval of 1-3 weeks With the indicated dose range, no adverse toxicological effects will be observed with the compounds of the invention which would preclude their administration to suitable individuals Each reference disclosed herein is incoφorated by reference herein in its entirety Any patent application to which this application claims priority is also incoφorated by reference herein in its entirety GLOSSARY The following definitions are provided to facilitate understanding of certain terms used frequently herein "Dιsease(s)" means and disease caused by or related to infection by a virus, including heφes labia s. heφes encephalitis, keratitis "Host cell" is a cell which has been transformed, transfected or infected, or is capable of transformation, transfection or infection by an exogenous polynucleotide sequence or virus "Identity," as known in the art, is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as the case may be, as determined by compaπng the sequences In the art, "identity" also means the degree of sequence relatedness between polypeptide or polynucleotide sequences, as the case may be, as determined by the match between strings of such sequences "Identity" can be readily calculated by known methods, including but not limited to those described in (Computational Molecular Biology, Lesk, A M , ed , Oxford University Press, New York, 1988, Biocomputing Informatics and Genome Projects, Smith, D W , ed , Academic Press, New York, 1993, Computer Analysis of Sequence Data, Part I. Griffin, A M , and Griffin, H G., eds , Humana Press, New Jersey, 1994. Sequence Analysis in Molecular Biology, von Heinje, G Academic Press, 1987, and Sequence Analysis Primer, Gπbskov, M and Devereux, J , eds , M Stockton Press, New York, 1991 , and Caπllo, H , and Lipman, D , SIAM J Applied Math , 48 1073 (1988) Methods to determine identity are designed to give the largest match between the sequences tested Moreover, methods to determine identity are codified in publicly available computer programs Computer program methods to determine identity between two sequences include, but are not limited to, the GCG program package (Devereux, J , et al , Nucleic Acids Research 12(1) 387 (1984)), BLASTP, BLASTN, and FASTA (Atschul. S F et al , J Molec Biol 215 403-410 (1990) The BLAST X program is publicly available from NCBI and other sources (BLAST Manual, Altschul, S , et al , NCBI NLM NIH Bethesda, MD 20894, Altschul, ~ , et al , J Mol Biol 215 403-410 (1990) The well known Smith Waterman algorithm may also be used to determine identity Parameters for polypeptide sequence companson include the following 1) Algorithm Needleman and Wunsch, J Mol Biol 48 443-453 (1970) Comparison matrix BLOSSUM62 from Hentikoff and Hentikoff Proc Natl Acad Sci USA 89 10915-10919 (1992) Gap Penalty 12 Gap Length Penalty 4 A program useful with these parameters is publicly available as the gap program from Genetics Computer Group, Madison WI The aforementioned parameters are the default parameters for peptide comparisons (along with no penalty for end gaps) Parameters for polynucleotide comparison include the following 1) Algorithm Needleman and Wunsch, J Mol Biol 48 443-453 ( 1970) Comparison matrix matches = + 10, mismatch = 0 Gap Penalty 50 Gap Length Penalty 3 Available as The "gap" program from Genetics Computer Group, Madison WI These are the default parameters for nucleic acid comparisons A preferred meaning for "identity" for polynucleotides and polypeptides, as the case may be, are provided in (1) and (2) below (1) Polynucleotide embodiments further include an isolated polynucleotide comprising a polynucleotide sequence having at least a 50, 60, 70, 80, 85, 90, 95, 97 or 100% identity to the reference sequence of SEQ ID NO 1, wherein said polynucleotide sequence may be identical to the reference sequence of SEQ ID NO 1 or may include up to a certain integer number of nucleotide alterations as compared to the reference sequence, wherein said alterations are selected from the group consisting of at least one nucleotide deletion, substitution, including transition and transversion, or insertion, and wherein said alterations may occur at the 5' or 3' terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among the nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence, and wherein said number of nucleotide alterations is determined by multiplying the total number of nucleotides in SEQ ID NO 1 by the integer defining the percent identity divided by 100 and then subtracting that product from said total number of nucleotides in SEQ ID NO 1, or n n < x n - (x n • y). wherein n n is the number of nucleotide alterations, x n is the total number of nucleotides in SEQ ID NO 1 , y is 0 50 for 50%, 0 60 for 60%, 0 70 for 707c, 0 80 for 80%, 0 85 for 85%, 0 90 for 90%, 0 95 for 95%, 0 97 for 97% or 1 00 for 100%, and • is the symbol for the multiplication operator, and wherein any non-integer product of x n and y is rounded down to the nearest integer prior to subtracting it from x n Alterations of a polynucleotide sequence encoding the polypeptide of SEQ ID NO 2 may create nonsense, missense or frameshift mutations in this coding sequence and thereby alter the polypeptide encoded by the polynucleotide following such alterations By way of example, a polynucleotide sequence of the present invention may be identical to the reference sequence of SEQ ID NO 2, that is it may be 100% identical, or it may include up to a certain integer number of amino acid alterations as compared to the reference sequence such that the percent identity is less than 100% identity Such alterations are selected from the group consisting of at least one nucleic acid deletion, substitution, including transition and transversion, or insertion, and wherein said alterations may occur at the 5' or 3' terminal positions of the reference polynucleotide sequence or anywhere between those terminal positions, interspersed either individually among the nucleic acids in the reference sequence or in one or more contiguous groups within the reference sequence The number of nucleic acid alterations for a given percent identity is determined by multiplying the total number of amino acids in SEQ ID NO 2 by the integer defining the percent identity divided by 100 and then subtracting that product from said total number of amino acids in SEQ ID NO 2, or n n < x n - (x n • y). wherein n n is the number of amino acid alterations, x n is the total number of amino acids in SEQ ID NO 2, y is, for instance 0 70 for 70%, 0.80 for 80%, 0.85 for 85% etc., • is the symbol for the multiplication operator, and wherein any non-integer product of x n and y is rounded down to the nearest integer prior to subtracting it from x n (2) Polypeptide embodiments further include an isolated polypeptide comprising a polypeptide having at least a 50,60, 70, 80, 85, 90, 95, 97 or 100% identity to a polypeptide reference sequence of SEQ ID NO 2, wherein said polypeptide sequence may be identical to the reference sequence of SEQ ID NO. 2 or may include up to a certain integer number of amino acid alterations as compared to the reference sequence, wherein said alterations are selected from the group consisting of at least one amino acid deletion, substitution, including conservative and non-conservative substitution, or insertion, and wherein said alterations may occur at the amino- or carboxy-terminal positions of the reference polypeptide sequence or anywhere between those terminal positions, interspersed either individually among the amino acids in the reference sequence or in one or more contiguous groups within the reference sequence, and wherein said number of amino acid alterations is determined by multiplying the total number of amino acids in SEQ ID NO 2 by the integer defining the percent identity divided by 100 and then subtracting that product from said total number of amino acids in SEQ ID NO 2, or "a ≤ x a " ( χ a * )- wherein n a is the number of amino acid alterations, x a is the total number of ammo acids in SEQ ID NO.2, y is 0.50 for 50%, 0.60 for 60%, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, 0.90 for 90%, 0 95 for 95%, 0 97 for 97% or 1.00 for 100%, and • is the symbol for the multiplication operator, and wherein any non-integer product of x a and y is rounded down to the nearest integer prior to subtracting it from x a By way of example, a polypeptide sequence of the present invention may be identical to the reference sequence of SEQ ID NO.2, that is it may be 100% identical, or it may include up to a certain integer number of amino acid alterations as compared to the reference sequence such that the percent identity is less than 100% identity Such alterations are selected from the group consisting of at least one amino acid deletion, substitution, including conservative and non-conservative substitution, or insertion, and wherein said alterations may occur at the amino- or carboxy-terminal positions of the reference polypeptide sequence or anywhere between those terminal positions, interspersed either individually among the amino acids in the reference sequence or in one or more contiguous groups within the reference sequence The number of amino acid alterations for a given % identity is determined by multiplying the total number of amino acids in SEQ ID NO 2 by the integer defining the percent identity divided by 100 and then subtracting that product from said total number of amino acids in SEQ ID NO 2, or n a < x a - (x a • y), wherein n a is the number of amino acid alterations. x a is the total number of amino acids in SEQ ID NO 2, y is, for instance 0 70 for 70%, 0 80 for 80%, 0 85 for 85% etc , and • is the symbol for the multiplication operator, and wherein any non-integer product of x a and y is rounded down to the nearest integer prior to subtracting it from x a "Isolated" means altered "by the hand of man" from its natural state, i e , if it occurs in nature, it has been changed or removed from its onginal environment, or both For example, a polynucleotide or a polypeptide naturally present in a living organism is not "isolated," but the same polynucleotide or polypeptide separated from the coexisting matenals of its natural state is "isolated" as the term is employed herein Moreover, a polynucleotide or polypeptide that is introduced into an organism by transformation, genetic manipulation or by any other recombinant method is ' isolated" even if it is still present in said organism, which organism may be living or non-living "Polynucleotιde(s)" generally refers to any polynbonucleotide or polydeoxπbonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA "Polynucleotιde(s)" include, without limitation, single- and double- stranded DNA, DNA that is a mixture of single- and double-stranded regions or single-, double- and tnple-stranded regions, single- and double-stranded RNA, and RNA that is mixture of single- and double- stranded regions, hybrid molecules compnsing DNA and RNA that may be single-stranded or, more typically, double-stranded, or tnple-stranded regions, or a mixture of single- and double- stranded regions In addition, polynucleotide" as used herein refers to tnple-stranded regions compnsing RNA or DNA or both RNA and DNA The strands in such regions may be from the same molecule or from different molecules The regions may include all of one or more of the molecules, but more typically involve only a region of some of the molecules One of the molecules of a tnple-hehcal region often is an ohgonucleotide As used herein, the term "polynucleotιde(s)" also includes DNAs or RNAs as descnbed above that contain one or more modified bases Thus, DNAs or RNAs with backbones modified for stability or for other reasons are "polynucleotιde(s) ' as that term is intended herein Moreover, DNAs or RNAs compnsing unusual bases, such as inosine, or modified bases, such as tntylated bases, to name just two examples, are polynucleotides as the term is used herein It will be appreciated that a great vanety of modifications have been made to DNA and RNA that serve many useful puφoses known to those of skill in the art The term "polynucleotιde(s)" as it is employed herein embraces such chemically, enzymatically or metabolically modified forms of polynucleotides, as well as the chemical forms of DNA and RNA charactenstic of viruses and cells, including, for example, simple and complex cells "Polynucleotιde(s)" also embraces short polynucleotides often referred to as ohgonucleotιde(s) "Polypeptιde(s)" refers to any peptide or protein compnsing two or more ammo acids joined to each other by peptide bonds or modified peptide bonds "Polypeptιde(s)" refers to both short chains, commonly referred to as peptides, o gopeptides and o gomers and to longer chains generally referred to as proteins Polypeptides may contain amino acids other than the 20 gene encoded amino acids "Polypeptιde(s)" include those modified either by natural processes, such as processing and other post-translational modifications, but also by chemical modification techniques Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature, and they are well known to those of skill in the art It will be appreciated that the same type of modification may be present the same or varying degree at several sites in a given polypeptide Also, a given polypeptide may contain many types of modifications Modifications can occur anywhere m a polypeptide. including the peptide backbone, the amino acid side-chains, and the amino or carboxyl termini Modifications include, for example, acetylation, acylation, ADP-nbosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide denvative, covalent attachment of a pid or hpid denvative, covalent attachment of phosphotidyhnositol, cross-linking, cychzation, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteme, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, lodination, methylation, mynstoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, glycosylation, hpid attachment, sulfation, gamma-carboxylation of glutamic acid residues, hydroxylation and ADP-nbosylation, selenoylation, sulfation, transfer-RNA mediated addition of ammo acids to proteins, such as argmylation, and ubiquitination See, for instance, PROTEINS - STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed , T E Creighton, W H Freeman and Company, New York (1993) and Wold, F , Posttranslational Protein Modifications Perspectives and Prospects, pgs 1-12 in POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B C Johnson, Ed , Academic Press, New York ( 1983), Seifter et al , Meth Enzymol 182 626-646 (1990) and Rattan et al , Protein Synthesis Posttranslational Modifications and Aging, Ann N Y Acad Sci 663 48-62 ( 1992) Polypeptides may be branched or cyclic, with or without branching Cyclic, branched and branched circular polypeptides may result from post-translational natural processes and may be made by entirely synthetic methods, as well "Vanant(s)" as the term is used herein, is a polynucleotide or polypeptide that differs from a reference polynucleotide or polypeptide respectively, but retains essential properties A typical vanant of a polynucleotide differs in nucleotide sequence from another, reference polynucleotide Changes in the nucleotide sequence of the vanant may or may not alter the amino acid sequence of a polypeptide encoded by the reference polynucleotide Nucleotide changes may result in amino acid substitutions, additions, deletions, fusions and truncations in the polypeptide encoded by the reference sequence, as discussed below A typical variant of a polypeptide differs in amino acid sequence from another, reference polypeptide Generally, differences are limited so that the sequences of the reference polypeptide and the vanant are closely similar overall and, in many regions, identical A vanant and reference polypeptide may differ in amino acid sequence by one or more substitutions, additions, deletions in any combination A substituted or inserted amino acid residue may or may not be one encoded by the genetic code A vanant of a polynucleotide or polypeptide may be a naturally occurring such as an allelic variant, or it may be a variant that is not known to occur naturally Non-naturally occurring variants of polynucleotides and polypeptides may be made by mutagenesis techniques, by direct synthesis, and by other recombinant methods known to skilled artisans EXAMPLES The examples below are earned out using standard techniques, which are well known and routine to those of skill in the art, except where otherwise descnbed in detail The examples are illustrative, but do not limit the invention The identification of gene expression following infection with HSV-1 Tissue culture cells are infected with HSV- 1 strain KOS and harvested post- mfection for RNA isolation HSV- 1 infected tissue culture cells are efficiently disrupted and processed in the presence of chaotropic agents and RNase inhibitor to provide a mixture of cellular and viral RNAs RNA may then be polyA+ selected for use in Northern blotting, with hybridization of a RNA probe specific to a region within the HSV- 1 genome RNase free, DNase free, DNA and protein free preparations of total or polyA-i- selected RNA are suitable for 5' end mapping of the gene of interest, using for example techniques such as 5' RACE (rapid amplification of cDNA ends, Gibco-BRL) Unique pnmer pairs for Reverse Transcription PCR (RT-PCR), used in the 5' RACE procedure, are designed from the published sequence of HSV- 1 strain 17, which is homologous to that of HSV- 1 strain KOS Example 1 RNA isolation, Northern blotting and 5' end identification The polynucleotide having a DNA sequence given in Table 1 [SEQ ID NO 1 ] was identified from a Northern blot of polyA+ RNA harvested from host cells infected with HSV-1 strain KOS, using a complementary radiolabelled fragment of RNA as probe prepared by routine methods, for example a) Viral infection and RNA isolation Tissue culture cells such as Vero (monkey kidney cells) are seeded into 850 cm^ tissue culture roller bottles in tissue culture medium [Dulbecco s Modified Eagle Medium (DMEM, Gibco BRL, Life Technologies) supplemented with 10% heat inactivated fetal bovine serum (FBS. HyClone Laboratories, Inc ), 1 % penicillin-streptomycin (Gibco BRL, Life Technologies), 5 ml per liter of 200 mM L-glutamine (Gibco BRL, Life Technologies), 10 ml per liter of 7 5% (w/v) sodium bicarbonate solution (Gibco BRL, Life Technologies)] and incubated at 37°C Cells are infected by addition of HSV- 1 (strain KOS) in a low volume of tissue culture medium at a multiplicity of infection of 10 plaque forming units per cell After a 1 hour infection at 37°C, additional DMEM is added and infected cells are harvested at 17 hours post-infection For total RNA isolation TRI Reagent® (Molecular Research Center, Inc) is used according to the manufacturer s instructions In a biological safety cabinet infected tissue culture cells are harvested by removal of tissue culture medium and the addition of 6 7 ml of TRI Reagent® per roller bottle ( 1 ml per 75 mg of cells) The mixture is then vortexed until homogenous Typically, two 850 cm^ roller bottles are used per isolation, each containing approximately 500 mg of cells per roller bottle The homogenate is centnfuged at 12.000 x g for 10 min at 4°C to remove the insoluble material and the supernatant is transferred to a fresh tube and allowed to stand for 5 min at room temperature To this mixture 1 34 ml of BCP Phase Separation Reagent is added (Molecular Research Center, Inc, 0 2 ml per ml of TRI Reagent®), vortexed vigorously for 15 seconds and allow to stand for 5 min at room temperature This mixture is then centnfuged at 12000 x g for 20- 30 min at 4°C Approximately 75% of the colorless upper aqueous phase (containing RNA) is transferred to fresh tubes and 3 35ml of isopropanol is added (0.5ml of isopropanol per ml of TRI Reagent) The samples are vortexed and allowed to stand for 5 min at room temperature Samples are frozen in a dry ice / ethanol bath until solid and centnfuged at 12,000 x g for 30 min at 4°C The supernatant is removed and the RNA pellet is air dned for 10 minutes at room temperature and resuspended in 503 ul of TNM buffer (75 ul of TNM buffer per lOOmg of starting material, TNM 40 mM Tns-HCl pH 8 0, 10 mM NaCl, 6 mM MgCl 2 ). DNA is removed by the addition of 75 units of RNase-free DNase 1 (Gibco BRL, Life Technologies) per 100 ul of RNA and incubated at 37°C for 20 mm To this is added 10% of sample volume of 4M LiCl The samples are transferred to eppendorf tubes and extracted with an equal volume of phenol chloroform ( 1 1 v v) The samples are centnfuged for 3 min at 13,000 φm in an eppendorf centπfuge, the aqueous phase is transferred to a clean tube and extracted with an equal volume chloroform Again the aqueous phase is transferred to a new tube and double the sample volume of ethanol is added The samples are frozen until solid in dry ice / ethanol bath and centnfuged at 13,000 φm in an eppendorf centrifuge for 30 mm at 4°C The ethanol is removed and the RNA is washed with 75% ethanol (minimum 1 ml), vortexed and the RNA pelleted by centnfugation in an eppendorf centrifuge at 13,000 φm for 5 min at 4°C The RNA pellet is air dried for 10 mm and resuspended in 200 ul TNM buffer The concentration of RNA is determined by measuring the absorbance at 260 nm Fifty micrograms of RNA is digested a second time using 10 units of DNase I (RNase and protease free, Wortrnngton Biochemical Coφ ) and incubated at 37°C for 30 min, followed by addition of 4M LiCl, extraction, precipitation and quantitation as described above. The quality of the RNA is assessed by electrophoresis of 1 ug in a 1% agarose gel and visualized following staining with ethidmm bromide [Sambrook et al., MOLECULAR CLONING, A LABORATORY MANUAL, (supra)] For polyA+ selected RNA the Poly(A)Pure mRNA isolation kit (Ambion) is used according to the manufacturer ' s instructions In a biological safety cabinet infected tissue culture cells are harvested by removal of tissue culture medium and addition of 10 ml of Lysis solution per 2 x 10 8 cells Typically, three 850 cm 2 roller bottles are used per isolation, each containing approximately 4 6 x 10^ cells The lysates are combined, transferred to a 50 ml tube and vortexed until homogenous Two starting volumes (20 ml) of Dilution Buffer are added to the lysate and mixed by inversion The lysate is centnfuged at 12,000 x g at 4°C for 15 minutes to remove particulate cellular debris and the supernatant is transferred to a clean 50 ml conical tube One pre-packaged vial of o go dT cellulose is added to the lysate and mixed by inversion The tube is rocked gently for 60 minutes at room temperature and the o go dT resin is pelleted by centnfugation at 2,000 x g for 3 minutes at room temperature The supernatant is removed and the resin washed by addition of 10 ml of Binding Buffer, mixed well by inversion, and pelleting the resin as before This wash is repeated twice more using 10 ml of Binding Buffer The resin is then washed by resuspension in 10 ml of Wash Buffer, mixed well by inversion, and pelleting the resin as before This wash is repeated twice more using 10 ml of Wash Buffer After the third wash, the ohgo dT resin is resuspended in 1 ml of Wash Buffer and transferred to a spin column placed in a stenle microfuge tube The spin column is centnfuged at 5,000 x g at room temperature for 10 seconds, and the absorbance of the flowthrough is measured at 260 nm to ensure that it is less than 0 05 The spin column is placed into a new microfuge tube and the polyA+ RNA is eluted by addition of 200 μl of prewarmed (65°C) Elution Buffer and centnfugation at 5,000 x g at room temperature for 30 seconds The elution is repeated with an additional 200 μl of Elution Buffer and the polyA+ RNA is precipitated by addition of 40 μl of 5M sodium acetate, 2 μl of glycogen, and 2 5 volumes of 100% ethanol The RNA is stored at -80°C for 30 minutes and recovered by centnfugation at 12,000 x g for 20 minutes at 4°C The supernatant is removed and the RNA pellet is air dried for 10 minutes at room temperature and resuspended in 40 μl of DEPC-treated water/0 1 mM EDTA The concentration of RNA is determined by measuring the absorbance at 260 nm, the RNA preparations are stored at -80°C until use [Sambrook et al , MOLECULAR CLONING, A LABORATORY MANUAL, (supra)] b) Northern blotting of HSV-1 polyA+ selected RNA to identify novel transcripts PolyA+ selected RNA (5 ug) isolated from HSV- 1 (KOS) infected Vero cells at 17 hours post-mfection, or from uninfected (mock) Vero cells, are run on a 1 % agarose gel prepared using 10X Denatunng Gel Buffer (Ambion) Each RNA sample is suspended in 10 μl of DEPC-treated water and three volumes (30 μl) of Sample Loading Dye (Ambion) Samples are heated to 65°C prior to loading onto the gel to denature RNA secondary structure Gels are run in IX Gel Running Buffer (Ambion) at 65 volts for 3 to 4 hours After electrophoresis the RNA is transferred to a positively charged nylon membrane (BnghtStar-Plus, Ambion) by vacuum transfer at 5 inches Hg for 90 minutes in 5X SSC, 10 mM NaOH After the transfer, the membrane is rinsed briefly IX Gel Running Buffer and is UV crosshnked (Stratagene) Cross nked RNA membranes can be stored at -20°C in a vessel that will protect them from physical damage Riboprobes are generated from plasmid DNA specific for the UL8/UL9 region of the HSV genome (nboprobe 8R-3 . K Baradaran, C E Dabrowski, and P A Schaffer, 1994 J Virol 68 4251-4261) using the Riboprobe In Vitio Transcription System (Promega) p8R-3' plasmid DNA is linearized with Xbal and 2 ug is transcribed and radiolabelled according to the manufacturer s instructions Riboprobes are purified by application to ProbeQuant G-50 Micro Columns (Pharmacia) followed by centnfugation at 3000 φm for two minutes in the Eppendorf microcentnfuge, and repeated to remove free nucleotides The membrane to which the RNA is bound is prehybndized for 1 hour in 10 ml Prehybndization/Hybridization Solution (Ambion) per 100 cm 2 of membrane at 75°C The prehybndization solution is removed and replaced with 10 ml Hybridization solution consisting of Prehybndization/Hybridization Solution preheated to the hybridization temperature (75°C ) plus 1 x 10^ cpm per ml of the 8R-3' riboprobe The membrane is hybridized overnight at 75°C and then washed twice with Low Stringency Wash Solution (Ambion) for 5 minutes each at room temperature, three times with High Stringency Wash Solution (Ambion) for 20 minutes each at 75 C C and autoradiographed c) The rapid amplification of 5' cDNA ends from RNA samples derived from infected cells Total RNA ( 1 ug) is reverse transcribed using the 5' RACE System for Rapid Amplification of cDNA Ends (Gibco BRL. Life Technologies) according to the manufacturer s instructions for first strand cDNA synthesis of transcripts with high GC content Gene specific primer 1 ( 100 nM GSP1 , 5'-GCA GAT GTT ATC GCC-3') is used to prime each reaction Controls without the addition of Superscript II reverse transcπptase are included All samples are then treated with RNase mix and the cDNA is punfied using the GlassMax DNA Isolation System included with the 5' RACE kit Following purification. cDNA samples are tailed with dCTP using terminal deoxynucleotidyl transferase (TdT) and used in a PCR reaction using an Abridged Anchor Pnmer (supplied with the 5' RACE kit) and a second gene specific primer (GSP2, 5'-GAT GCA GAC GTT ATC TC-3') PCR reactions are assembled on ice in 0 2 ml thin-wall tubes, according to the instructions for PCR of dC-tailed cDNA Reaction tubes are transferred directly from ice to the thermal cycler preequi brated to the initial denaturation temperature (94°C) PCR reactions are run on a Perkin Elmer GeneAmp PCR System 9600 as follows 2 minutes at 94 C, then 35 cycles of 30 seconds at 94 C 30 seconds at 55 C and 1 minute, 30 seconds at 72 C followed by 5 minutes at 72 C and then a hold temperature of 5 C Aliquots ( 10 ul) are then electrophoresed on 6% acrylamide gels in IX TBE (Novex) and stained with ethidium bromide The size of the PCR product is estimated by comparison to a 100 bp DNA Ladder (Gibco BRL, Life Technologies) A nested PCR amplification is then performed following the instructions for Nested Amplification using the Universal Amplification Primer (UAP) and a third gene specific pnmer (GSP3, 5'-CAU CAU CAU CAU CCG ACC ACC ACA TG-3') PCR is performed as descnbed above The primers used in the nested amplification contain sequences that are specific for cloning the PCR product using the CloneAmp pAMPl System (Gibco BRL, Life Technologies) The PCR products are purified from a 6% acrylamide gel and cloned into the pAMPl vector following the manufacturer s instructions After the PCR product is annealed to the pAMPl vector the reaction mixture is transformed into DH5α cells (Gibco BRL, Life Technologies) following standard protocols [Sambrook et al , MOLECULAR CLONING A LABORATORY MANUAL, (supra)] Bacteria are plated on LB plates containing 100 ug/ml ampicilhn (Digene Diagnostics Inc ), grown overnight at 37°C and colonies are picked for DNA minipreps Plasmid DNA is isolated from 5 ml bacterial cultures using the RPM Kit (Bio 101 ) following the manufacturer s instructions Plasmid DNA is analyzed by restriction digest with BamHI and EcoRI and electrophoresed on 6% acrylamide gels in IX TBE (Novex) to determine the size of the insert compared to a 100 bp DNA ladder as above Clones are then used in sequencing reactions (T7 Sequenase 7-deaza-dGTP Sequencing Kit Amersham Life Science) with the SP6 promoter primer (Gibco BRL. Life Technologies) to determine the 5 end of the cDNA The 5' end is determined following sequencing of DNA from two or more clones containing amplified PCR products and used to identify the contiguous DNA sequence in SEQ ID NO 1 Example 2 OBP-NT Characterization A novel messenger RNA (mRNA) encoding the newly described protein. OBP-NT, was identified by Northern blotting utilizing an RNA probe complementary to the UL8/9 region of the HSV-1 genome (see example 1) While previous efforts to map RNAs encoded within this region have been descnbed (Deb, S P et al 1993 Biochem and Biophys Res Comm 193 617-623, Baradaran, K et al 1994 J Virol 68 4251-4261), the current study utilized mRNA isolated at a very late time-point ( 17 hours post-infection) Identification of the 5 end of the mRNA, as described above, together with knowledge of the 3' end from previous mapping studies (Baradaran, K et al 1994 J Virol 68 4251- 4261), allowed for the prediction of the OBP-NT open reading frame according to well- established guidelines in eukaryotes (Kozak, M 1986 Cell 44 283-292) a) Predicted OBP-NT features From this analysis OBP-NT is predicted to encode a protein in-frame within the well- characterized viral origin binding protein, OBP, with the lnititiatmg methiomne of OBP-NT corresponding to amino acid 142 of OBP Thus, OBP-NT shares with OBP a dimenzation domain (leucine zipper at amino acids 150-171 , Hazuda, D J et al 1992 J Biol Chem 267 14309-14315), helicase motifs II through VI (Malik, A K and S K Weller 1996 J Virol 70 7859-7866), and the ongin specific DNA binding domain at the C-temimus (Deb, S and S W Deb 1991 J Virol 65 2829-2838) Based on these shared sequences between the proteins, OBP-NT is expected to be able to form homodimers with itself and heterodimers with OBP, through the conserved leucine zipper region at amino acids 150- 171 (OBP numbering) These proteins may also be able to dimeπze with as yet undefined viral or cellular proteins OBP-NT is not expected to function as a helicase as a monomer or homodimer although it retains conserved helicase motifs II through VI, as the missing motifs I and la are critical for nucleotide binding and thus helicase activity However, OBP-NT may function as a helicase as a heterodimer with, for example, OBP, and as such may have similar or different properties as compared to the OBP homodimer OBP-NT is also predicted to bind sequences within the viral ongins of replication, expeπments demonstrating this ability are described below b) OBP-NT binding to viral origin of replication sequences Evidence for the ability of OBP-NT to bind to sequences within the viral origins of replication (onS and onL) can be found in a number of published sources, in results from experiments performed to charactenze origin binding by the other members of the OBP family, OBP and OBPC For example, electrophoretic mobility shift assays (EMSA) experiments have been published in which extracts from HSV- 1 infected cells were incubated with radiolabeled onS DNAs (sites I, II or III, see figure 1) In these experiments two to three viral specific complexes can be identified - complex A, comprised of OBPC and site I or site II DNAs (Baradaran, K et al 1994 J Virol 68 4251-4261), and complex B, compnsed of OBP and site I, II or III DNAs, which appears as a single band or as a doublet (see Dabrowski, C E and P A Schaffer 1991 J Virol 65 3140-3150, Weir, H M et al 1989 Nucl Acids Res 17 1409-1425, Hardwicke, M A and P A Schaffer 1995 J Virol 69 1377-1388) In the presence of antibody directed against the C- terminal ammo acids of OBP (which are also in common with OBP-NT and OBPC), complex A is shifted to a higher molecular weight complex (supershifted). The ρrotein:DNA complexes identified as complex B formed with oriS DNAs (sites I, II, III) also shifted or significantly decreased in intensity in the presence of antibody directed against the C-terminus of OBP (Figure 8, Dabrowski, C. E. and P. A. Schaffer. 1991. J. Virol. 65:3140-3150). However, as shown in Figure 10 in this paper, complex B formed with extracts from HSV infected cells appears as a single band in these experiments, and demonstrates a different electrophoretic mobility than the complex (designated 9-1) formed with partially purified recombinent OBP. The resulting difference in electrophoretic mobilities between the 'complex B' and 'complex 9- 1 ' proteins, and the ability of these proteins to be supershifted in EMSA experiments by antibody directed against the C-terminus of OBP, is indicative of the presence of OBP-related proteins of slightly different molecular weights. This result is consistent with the presence of full-length OBP within complex 9-1, formed with partially purified OBP, and a slightly smaller molecular weight protein translated in-frame with OBP, i.e. OBP-NT, within complex B from infected cell extracts. Thus, by EMSA. complex B is observed to be one or two bands in complexes formed from HSV infected cell extracts and oriS site I or II DNAs, while in contrast, complex B appears to be a single band with site III as probe; no complex B doublet has been observed (Dabrowski, C. E. and P. A. Schaffer. 1991. J. Virol. 65:3140-3150; Weir, H. M. et al. 1989. Nucl. Acids Res. 17: 1409-1425; Hardwicke, M. A. and P. A. Schaffer. 1995. J. Virol. 69: 1377-1388); the lower band of the doublet is now believed to be comprised of OBP-NT and the upper band comprised of OBP. The presence of one or both bands appears to be dependent on the extract preparation, time of harvest post-infection, and length of exposure to film which may blur the presence of 2 discrete bands. Further evidence for the ability of OBP-NT to bind sequences within the viral origins of replication are shown in figures 3 and 4, following EMSA analysis of extracts from HSV-1 infected cells incubated with radiolabeled oriS DNAs. For these experiments Vero cells are seeded into 850 cm 2 tissue culture roller bottles in DMEM containing 10% FBS and incubated overnight at 37°C. Cells are either mock infected or infected with HSV-1 (KOS) at an MOI of 10 and harvested at 3, 6, 9, 12, 15, or 18 hours post infection (hpi). Total cell extracts are prepared by pelleting the cells at 2000 φm at 4°C for 5 minutes in a tabletop centrifuge, washing the pellet twice with phosphate buffered saline (PBS; Sambrook et al, MOLECULAR CLONING, A LABORATORY MANUAL) and resuspending in 50 ul NET buffer (50 mM Tris, pH 8, 100 mM NaCl, 1 mM EDTA). Pellets are frozen at -80°C, thawed at 37°C and sonicated twice for 30 seconds each on a setting of 4 using a Virsonic 550 Ultrasonic Cell Disrupter (VirTis). The cell debris is pelleted at 14000 φm in an Eppendorf microcentrifuge for 10 minutes at 4°C, and the supernatant is aliquoted and stored at -80°C. Protein quantitation is performed on the total cell extracts using the Bio-Rad Protein Assay (Bio-Rad) following the_ manufacturer's instructions for the Microassay Procedure. Radiolabeled HSV- 1 onS sites I, II, III and III(I) probes are prepared as previously described (Dabrowski, CE. and P. A Schaffer. 1991. J Virol. 65 3140-3150). The site III(I) probe contains sequences identical to site III with a central divergent nucleotide (T) changed to a G, as found in site I For the EMS As, mock infected (10 ug protein) or HSV infected (2 ug protein) cell extracts are incubated with 2 ng of probe and 1.5 ug of poly(dl-dC) poly(dl-dC) (Pharmacia) in DNA binding buffer (10% glycerol, 50 mM HEPES, pH 7.5 [N-2-hydroxyelthylpιperazιne-N'-2-ethanesulfonιc acid; Life Technologies], 0.1 mM EDTA, 0.5 mM DTT, 75 mM NaCl) in a final volume of 10 ul for 30 minutes at room temperature. Equal amounts of radioactivity are loaded onto each gel (35,000 cpm) and the protein DNA complexes are separated by electrophoresis in 6% polyacrylamide gels prepared and run in 0.5X TBE (Life Technologies) at 200V. Shown in figure 3 are the results of an EMSA performed with total cell extract harvested 12 hpi incubated with site I, II, III and III(I) probes Complex A containing OBPC (Baradaran, K. et al. 1994. J Virol. 68:4251-4261) is present with site I, II, and III(I) probes but not with the site III probe, as previously descnbed (Dabrowski, CE and P A. Schaffer. 1991. J Virol. 65.3140-3150). Complex B, a doublet of OBP (upper band of doublet) and OBP-NT (lower band of doublet), is present with site I, II, and III (I) probes, but only the lower OBP-NT band of the complex B doublet is present with the site III probe This experiment demonstrates that, despite the common C-terminal DNA binding region present in OBP, OBP-NT and OBPC, only OBP-NT is able to bind to site III within the core viral origin of replication Further, the inability of OBP and OBPC to bind site III is dependent on the single nucleotide difference between sites I and III within the core OBP binding domain (T in site III, G in site I), and now explains the difference in results described above (see background) whereby punfied OBP was shown not to bind site III, although a band of the approximate correct size was identified from HSV infected cell extracts EMSAs were also performed with radiolabeled onS probes using mock or HSV infected cell extracts harvested at increasing times post infection. As shown in figure 4A and B, with site I and II DNAs the complex B doublet appears by 6 hpi and increases in intensity through 18 hpi. Additional proteιn:DNA complexes (C and D) with decreased electrophoretic mobility are also seen at increasing times post infection, with the concomitant disappearance of the mock infected band. While the complex B doublet bands are of approximately equal intensity with the site I probe, the lower OBP-NT complex B band and the complex D band predominate at all times post-infection with the site II probe In EMSAs performed with the site III probe (figure 4C) complex B appears as a single discrete band, appearing at 9 hpi and increasing in intensity up to 18 hpi With this probe, complex D but not complex C is also formed; from these expeπments complex C appears to be OBP-related, whereas complex D appears to be OBP-NT related In addition, a lower light intensity band is apparent, which only increases significantly in intensity at 18 hpi; this band appears to be of slightly faster mobility than complex A formed with OBPC. Aligning the site I, II and III EMSAs, using the mock infected bands for reference, it becomes evident that the single band of complex B formed with the site III probe has an electrophoretic mobility identical to the lower band of the complex B doublet seen with the site I probe. From these experiments it we conclude that OBP and OBPC binds to sites I and II in the viral origins of replication, as previously reported. OBP-NT is also able to bind to sites I and II, and further, is the only viral protein of the known OBP family able to bind to site III. c) OBP-NT Expression in HSV-1 Infected Cells OBP-NT protein is identified in HSV infected cells by Western blot, using a polyclonal anti-peptide antibody raised against a 10 amino acid peptide from the N-terminus of OBP-NT (MNDRPFHRL), and protein extracts from uninfected (mock) Vero cells, Vero cells harvested at 6 hours post infection (hpi), or Vero cells harvested at 18 hpi with HSV-1. Vero cells are seeded at 2.5 x 10 6 cells per 100 mm dish in DMEM containing 10% FBS and incubated overnight at 37°C Cells are either mock-infected or infected with HSV-1 (KOS) at an MOI of 10. At 6 or 18 hpi cells are scraped into the medium and pelleted at 1500 φm for 10 minutes in a tabletop centrifuge. Each pellet is resuspended in 400 ul phosphate buffered saline (PBS; Sambrook et al, MOLECULAR CLONING, A LABORATORY MANUAL); 200 ul of lysis buffer (30 mM Tris, pH 8.0, 15 mM EDTA, 3% SDS) is added and the lysates are incubated on ice for 10 minutes. The lysates are sonicated twice for 30 seconds, and 5 volumes of acetone are added. The precipitates are stored at -20°C for 30 minutes and pelleted for 15 minutes at 4°C in an Eppendorf microcentrifuge at 14000 φm. The pellets are allowed to air dry, resuspended in 400 ul of SDS-PAGE loading buffer (NOVEX) containing 0.05% •-mercapotoethanol, and 40 ul each of the mock, 6 hpi and 18 hpi extracts is loaded onto an 8% SDS-PAGE gel, along with 10 ul (13 ug) of a crude extract containing recombinant OBP (rOBP; gift of S. Lee and R. Lehman, Stanford University). Proteins are separated by electrophoresis overnight at 9 mA/gel and transferred to a PVDF membrane (PolyScreen; NEN Research Products) for 1 hr at 24 V using a GENIE electrophoretic transfer apparatus (Idea Scientific Company). For Western blot analysis, the membrane is incubated in blocking solution [5% skim milk in TBS (10 mM Tris, pH 8.0, 150 mM NaCl)] for 1 hr at room temperature. After a quick wash in TBS, the blot is incubated with the primary antibody diluted 1 :250 in TBS containing 0.05% Tween 20 (TBST) for 1 hr at room temperature. The blot is then washed three times for 5 min each with TBST, incubated with the secondary antibody (goat anti- rabbit alkaline phosphatase conjugated antibody, IgG; Promega) diluted 1 :4000 in TBST for 30 min at room temperature, and washed three times for 5 min each with TBS. The substrate is prepared by adding 66 ul of NBT (Promega) to 10 ml of alkaline phosphatase substrate buffer (100 mM Tris, pH 8.0, 100 mM NaCl, 5 mM MgCl 2 ), followed by the addition of 33 ul of BCIP (Promega). The substrate is added to the blot, and color development is followed. The blot is washed twice with deionized water to stop the reaction. The results of the Western blot (Figure 5) show a faint protein band of the approximate expected size (78 kDa by calculation) in the 6 hpi infected cell extract lane (6h), and a heavy protein band of the same size is evident in the 18 hpi infected cell extract lane (18h); this protein is not present in mock infected cell extracts (M). This anti-peptide antibody also identified OBP from the recombinant baculovirus extract, but did not identify OBP from infected cell extracts, suggesting the possibility of a difference in protein configuration or modification between these sources Example 3 Herpes Viral in vitro replication Seven HSV-1 proteins have been identified as nesessary and sufficient for replication of plasmid DNA containing a viral origin of replication in the context of a mammalian or insect cell, by transient replication assays (Stow, N. D. 1992. J. Gen. Virol. 73:313-321; Boehmer, P. E. and Lehman, I. R. 1997. Annu. Rev. Biochem. 66:347-384). However, neither these seven purified proteins, nor extracts of these proteins from recombinant baculovirus infected cells, are sufficient for origin-directed replication in vitro, suggesting that additional proteins are required (Skaliter, R. and Lehman, I. R. 1994. Proc. Natl. Acad. Sci. USA 91:10665-10669; Boehmer, P. E. and Lehman, I. R. 1997. Annu. Rev. Biochem. 66:347-384). The OBP-NT protein has now been identified as a viral origin binding protein capable of binding to the sequences defined as sites I, II and in in the viral origins of replication; these binding sites are contained within the core origin regions required for efficient replication. Further, OBP-NT is the only viral origin binding protein which binds to an oligonucleotide containing site in. Thus, OBP-NT is likely to supply the additional viral functionality required, in the presence of the seven previously identified replication proteins, for origin-directed replication in vitro. In addition to OBP-NT, it is possible that cellular proteins such as found to be required for non-origin directed replication, may be necessary for origin-directed replication (Skaliter, R. and Lehman, I. R. 1994. Proc. Natl. Acad. Sci. USA 91:10665-10669).

Claims: What is claimed is: I. An isolated polynucleotide comprising a polynucleotide having at least a 70% identity to a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:2. 2. An isolated polynucleotide comprising a polynucleotide having at least a 70% identity to a polynucleotide encoding the same polypeptide expressed by the OBP-NT gene contained in the Herpes simplex virus type 1 (HSV-1). 3. An isolated polynucleotide comprising a polynucleotide encoding a polypeptide comprising an amino acid sequence which is at least 70% identical to the amino acid sequence of SEQ ID NO:2. 4 The polynucleotide of Claim 1 comprising a sequence selected from SEQ ID NO:l, SEQ ID NO: 3 and SEQ ID NO: 5. 5. The polynucleotide of Claim 1 which encodes a polypeptide comprising the amino acid sequence selected from SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6. 6. A vector comprising the polynucleotide of Claim 1. 7. A host cell comprising the vector of Claim 6. 8. A process for producing a OBP-NT polypeptide or fragment comprising culturing a host cell of claim 7 under conditions sufficient for the production of said polypeptide or fragment. 9. A polypeptide comprising an amino acid sequence which is at least 70% identical to the amino acid sequence of SEQ ID NO:2. 10. An OBP-NT polypeptide comprising an amino acid sequence selected from SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6. I I. An antagonist which inhibits the activity, binding or expression of the polypeptide of Claim 10. 12. A method for the treatment of an individual in need of heφes anti-viral treatment comprising: administering to the individual a therapeutically effective amount of the polypeptide of Claim 11. 13. A process for diagnosing a disease related to expression or activity of the polypeptide of Claim 9 in an individual comprising: (a) analyzing for the presence or amount of said polypeptide in a sample derived from the individual. 14. A method for identifying compounds which interact with and inhibit or activate an activity of the polypeptide of Claim 9 comprising: contacting a composition compnsing the polypeptide with the compound to be screened under conditions to permit interaction between the compound and the polypeptide to assess the interaction of a compound, such interaction being associated with a second component capable of providing a detectable signal in response to the interaction of the polypeptide with the compound, and determining whether the compound interacts with and activates or inhibits an activity of the polypeptide by detecting the presence or absence of a signal generated from the interaction of the compound with the polypeptide 15 A method for inducing an immunological response in a mammal which comprises inoculating the mammal with an OBP-NT polypeptide of Claim 9, or a fragment or vanant thereof, adequate to produce antibody and/or T cell immune response to protect said animal from disease 16 A method of inducing immunological response in a mammal which compnses delivering a nucleic acid vector to direct expression of OBP-NT polypeptide of Claim 9, or fragment or a variant thereof, for expressing said OBP-NT polypeptide, or a fragment or a vanant thereof in vivo in order to induce an immunological response

Inventors: DABROWSKI-AMARAL CHRISTINE E, GARCIA JAVIER J, GORCZYCA MICHELE M, DABROWSKI-AMARAL, CHRISTINE, E., GARCIA, JAVIER, J., GORCZYCA, MICHELE, M.
Applicants: DABROWSKI AMARAL CHRISTINE E, GARCIA JAVIER J, GORCZYCA MICHELE M, SMITHKLINE BEECHAM CORP, DABROWSKI-AMARAL, CHRISTINE, E., GARCIA, JAVIER, J., GORCZYCA, MICHELE, M., SMITHKLINE BEECHAM CORPORATION
Application year: 1999
Main classification: C12Q1
IPC codes: A61K38, A61K39, A61P31, C07K14, C12N1, C12N5, C12N15, C12P21, G01N33