Abstract
The equine herpesvirus 1 (EHV-1) immediate-early (IE) phosphoprotein is essential for the activation of transcription from viral early and late promoters and regulates transcription from its own promoter. The EHV-1 EICP22 protein, a homolog of ICP22 of herpes simplex virus, increased the in vitro DNA binding activity of the IE protein for sequences in the IE, early, and late promoters. The EICP22 protein affected the rate as well as the extent of the IE protein binding to promoter DNA sequences. To study the DNA binding activity of the IE protein, Trp493, Gln495, Asn496, and Lys498 of the WLQN region, which is directly involved in DNA binding, were replaced with Ser (IEW493S), Glu (IEQ495E), Ile (IEN496I), and Glu (IEK498E), respectively. Gel shift assays revealed that the glutathione S-transferase (GST)-IEQ495E(407-615) and GST-IEK498E(407-615) proteins failed to bind to the IE promoter, indicating that the Gln and Lys residues are important for the DNA binding activity. In the presence of the GST-EICP22 protein, DNA binding activity of the GST-IEQ495E(407-615) protein was restored, suggesting that the EICP22 protein cooperates with the IE protein to regulate EHV-1 gene expression. Transient-transfection assays also showed that the EICP22 protein allowed the IEQ495E mutant to be functional as a transactivator. These results are unique and may represent an important role for the EICP22 protein in EHV-1 gene regulation.
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- Allen G. P., Bryans J. T. Molecular epizootiology, pathogenesis, and prophylaxis of equine herpesvirus-1 infections. Prog Vet Microbiol Immunol. 1986;2:78–144. [PubMed] [Google Scholar]
- Anderson K. P., Costa R. H., Holland L. E., Wagner E. K. Characterization of herpes simplex virus type 1 RNA present in the absence of de novo protein synthesis. J Virol. 1980 Apr;34(1):9–27. doi: 10.1128/jvi.34.1.9-27.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Breeden C. A., Yalamanchili R. R., Colle C. F., O'Callaghan D. J. Identification and transcriptional mapping of genes encoded at the IR/Us junction of equine herpesvirus type 1. Virology. 1992 Dec;191(2):649–660. doi: 10.1016/0042-6822(92)90240-p. [DOI] [PubMed] [Google Scholar]
- Caughman G. B., Lewis J. B., Smith R. H., Harty R. N., O'Callaghan D. J. Detection and intracellular localization of equine herpesvirus 1 IR1 and IR2 gene products by using monoclonal antibodies. J Virol. 1995 May;69(5):3024–3032. doi: 10.1128/jvi.69.5.3024-3032.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cheung A. K. DNA nucleotide sequence analysis of the immediate-early gene of pseudorabies virus. Nucleic Acids Res. 1989 Jun 26;17(12):4637–4646. doi: 10.1093/nar/17.12.4637. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cullinane A. A., Rixon F. J., Davison A. J. Characterization of the genome of equine herpesvirus 1 subtype 2. J Gen Virol. 1988 Jul;69(Pt 7):1575–1590. doi: 10.1099/0022-1317-69-7-1575. [DOI] [PubMed] [Google Scholar]
- Davison A. J., Scott J. E. The complete DNA sequence of varicella-zoster virus. J Gen Virol. 1986 Sep;67(Pt 9):1759–1816. doi: 10.1099/0022-1317-67-9-1759. [DOI] [PubMed] [Google Scholar]
- DeLuca N. A., McCarthy A. M., Schaffer P. A. Isolation and characterization of deletion mutants of herpes simplex virus type 1 in the gene encoding immediate-early regulatory protein ICP4. J Virol. 1985 Nov;56(2):558–570. doi: 10.1128/jvi.56.2.558-570.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Faber S. W., Wilcox K. W. Association of the herpes simplex virus regulatory protein ICP4 with specific nucleotide sequences in DNA. Nucleic Acids Res. 1986 Aug 11;14(15):6067–6083. doi: 10.1093/nar/14.15.6067. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Freeman M. J., Powell K. L. DNA-binding properties of a herpes simplex virus immediate early protein. J Virol. 1982 Dec;44(3):1084–1087. doi: 10.1128/jvi.44.3.1084-1087.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gray W. L., Baumann R. P., Robertson A. T., Caughman G. B., O'Callaghan D. J., Staczek J. Regulation of equine herpesvirus type 1 gene expression: characterization of immediate early, early, and late transcription. Virology. 1987 May;158(1):79–87. doi: 10.1016/0042-6822(87)90240-6. [DOI] [PubMed] [Google Scholar]
- Gray W. L., Baumann R. P., Robertson A. T., O'Callaghan D. J., Staczek J. Characterization and mapping of equine herpesvirus type 1 immediate early, early, and late transcripts. Virus Res. 1987 Sep;8(3):233–244. doi: 10.1016/0168-1702(87)90018-9. [DOI] [PubMed] [Google Scholar]
- Grundy F. J., Baumann R. P., O'Callaghan D. J. DNA sequence and comparative analyses of the equine herpesvirus type 1 immediate early gene. Virology. 1989 Sep;172(1):223–236. doi: 10.1016/0042-6822(89)90124-4. [DOI] [PubMed] [Google Scholar]
- Guan K. L., Dixon J. E. Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal Biochem. 1991 Feb 1;192(2):262–267. doi: 10.1016/0003-2697(91)90534-z. [DOI] [PubMed] [Google Scholar]
- Harty R. N., O'Callaghan D. J. An early gene maps within and is 3' coterminal with the immediate-early gene of equine herpesvirus 1. J Virol. 1991 Jul;65(7):3829–3838. doi: 10.1128/jvi.65.7.3829-3838.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Henry B. E., Robinson R. A., Dauenhauer S. A., Atherton S. S., Hayward G. S., O'Callaghan D. J. Structure of the genome of equine herpesvirus type 1. Virology. 1981 Nov;115(1):97–114. doi: 10.1016/0042-6822(81)90092-1. [DOI] [PubMed] [Google Scholar]
- Holden V. R., Caughman G. B., Zhao Y., Harty R. N., O'Callaghan D. J. Identification and characterization of the ICP22 protein of equine herpesvirus 1. J Virol. 1994 Jul;68(7):4329–4340. doi: 10.1128/jvi.68.7.4329-4340.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holden V. R., Harty R. N., Yalamanchili R. R., O'Callaghan D. J. The IR3 gene of equine herpesvirus type 1: a unique gene regulated by sequences within the intron of the immediate-early gene. DNA Seq. 1992;3(3):143–152. doi: 10.3109/10425179209034010. [DOI] [PubMed] [Google Scholar]
- Holden V. R., Yalamanchili R. R., Harty R. N., O'Callaghan D. J. ICP22 homolog of equine herpesvirus 1: expression from early and late promoters. J Virol. 1992 Feb;66(2):664–673. doi: 10.1128/jvi.66.2.664-673.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holden V. R., Yalamanchili R. R., Harty R. N., O'Callaghan D. J. Identification and characterization of an equine herpesvirus 1 late gene encoding a potential zinc finger. Virology. 1992 Jun;188(2):704–713. doi: 10.1016/0042-6822(92)90525-t. [DOI] [PubMed] [Google Scholar]
- Holden V. R., Zhao Y., Thompson Y., Caughman G. B., Smith R. H., O'Callaghan D. J. Characterization of the regulatory function of the ICP22 protein of equine herpesvirus type 1. Virology. 1995 Jul 10;210(2):273–282. doi: 10.1006/viro.1995.1344. [DOI] [PubMed] [Google Scholar]
- Honess R. W., Roizman B. Regulation of herpesvirus macromolecular synthesis. I. Cascade regulation of the synthesis of three groups of viral proteins. J Virol. 1974 Jul;14(1):8–19. doi: 10.1128/jvi.14.1.8-19.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones P. C., Hayward G. S., Roizman B. Anatomy of herpes simplex virus DNA VII. alpha-RNA is homologous to noncontiguous sites in both the L and S components of viral DNA. J Virol. 1977 Jan;21(1):268–276. doi: 10.1128/jvi.21.1.268-276.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kim S. K., Smith R. H., O'Callaghan D. J. Characterization of DNA binding properties of the immediate-early gene product of equine herpesvirus type 1. Virology. 1995 Oct 20;213(1):46–56. doi: 10.1006/viro.1995.1545. [DOI] [PubMed] [Google Scholar]
- Kim Y., Geiger J. H., Hahn S., Sigler P. B. Crystal structure of a yeast TBP/TATA-box complex. Nature. 1993 Oct 7;365(6446):512–520. doi: 10.1038/365512a0. [DOI] [PubMed] [Google Scholar]
- Lieberman P. M., Berk A. J. A mechanism for TAFs in transcriptional activation: activation domain enhancement of TFIID-TFIIA--promoter DNA complex formation. Genes Dev. 1994 May 1;8(9):995–1006. doi: 10.1101/gad.8.9.995. [DOI] [PubMed] [Google Scholar]
- Mackem S., Roizman B. Regulation of herpesvirus macromolecular synthesis: temporal order of transcription of alpha genes is not dependent on the stringency of inhibition of protein synthesis. J Virol. 1981 Oct;40(1):319–322. doi: 10.1128/jvi.40.1.319-322.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maguire H. F., Hoeffler J. P., Siddiqui A. HBV X protein alters the DNA binding specificity of CREB and ATF-2 by protein-protein interactions. Science. 1991 May 10;252(5007):842–844. doi: 10.1126/science.1827531. [DOI] [PubMed] [Google Scholar]
- Matsumura T., Smith R. H., O'Callaghan D. J. DNA sequence and transcriptional analyses of the region of the equine herpesvirus type 1 Kentucky A strain genome encoding glycoprotein C. Virology. 1993 Apr;193(2):910–923. doi: 10.1006/viro.1993.1200. [DOI] [PubMed] [Google Scholar]
- McGeoch D. J., Dolan A., Donald S., Brauer D. H. Complete DNA sequence of the short repeat region in the genome of herpes simplex virus type 1. Nucleic Acids Res. 1986 Feb 25;14(4):1727–1745. doi: 10.1093/nar/14.4.1727. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McGeoch D. J., Dolan A., Donald S., Rixon F. J. Sequence determination and genetic content of the short unique region in the genome of herpes simplex virus type 1. J Mol Biol. 1985 Jan 5;181(1):1–13. doi: 10.1016/0022-2836(85)90320-1. [DOI] [PubMed] [Google Scholar]
- Perini G., Wagner S., Green M. R. Recognition of bZIP proteins by the human T-cell leukaemia virus transactivator Tax. Nature. 1995 Aug 17;376(6541):602–605. doi: 10.1038/376602a0. [DOI] [PubMed] [Google Scholar]
- Poffenberger K. L., Raichlen P. E., Herman R. C. In vitro characterization of a herpes simplex virus type 1 ICP22 deletion mutant. Virus Genes. 1993 Jun;7(2):171–186. doi: 10.1007/BF01702397. [DOI] [PubMed] [Google Scholar]
- Post L. E., Roizman B. A generalized technique for deletion of specific genes in large genomes: alpha gene 22 of herpes simplex virus 1 is not essential for growth. Cell. 1981 Jul;25(1):227–232. doi: 10.1016/0092-8674(81)90247-6. [DOI] [PubMed] [Google Scholar]
- Preston V. G., Davison A. J., Marsden H. S., Timbury M. C., Subak-Sharpe J. H., Wilkie N. M. Recombinants between herpes simplex virus types 1 and 2: analyses of genome structures and expression of immediate early polypeptides. J Virol. 1978 Nov;28(2):499–517. doi: 10.1128/jvi.28.2.499-517.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Purves F. C., Ogle W. O., Roizman B. Processing of the herpes simplex virus regulatory protein alpha 22 mediated by the UL13 protein kinase determines the accumulation of a subset of alpha and gamma mRNAs and proteins in infected cells. Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6701–6705. doi: 10.1073/pnas.90.14.6701. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rice S. A., Long M. C., Lam V., Schaffer P. A., Spencer C. A. Herpes simplex virus immediate-early protein ICP22 is required for viral modification of host RNA polymerase II and establishment of the normal viral transcription program. J Virol. 1995 Sep;69(9):5550–5559. doi: 10.1128/jvi.69.9.5550-5559.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Robertson G. R., Whalley J. M. Evolution of the herpes thymidine kinase: identification and comparison of the equine herpesvirus 1 thymidine kinase gene reveals similarity to a cell-encoded thymidylate kinase. Nucleic Acids Res. 1988 Dec 9;16(23):11303–11317. doi: 10.1093/nar/16.23.11303. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ruyechan W. T., Dauenhauer S. A., O'Callaghan D. J. Electron microscopic study of equine herpesvirus type 1 DNA. J Virol. 1982 Apr;42(1):297–300. doi: 10.1128/jvi.42.1.297-300.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schwyzer M., Wirth U. V., Vogt B., Fraefel C. BICP22 of bovine herpesvirus 1 is encoded by a spliced 1.7 kb RNA which exhibits immediate early and late transcription kinetics. J Gen Virol. 1994 Jul;75(Pt 7):1703–1711. doi: 10.1099/0022-1317-75-7-1703. [DOI] [PubMed] [Google Scholar]
- Sears A. E., Halliburton I. W., Meignier B., Silver S., Roizman B. Herpes simplex virus 1 mutant deleted in the alpha 22 gene: growth and gene expression in permissive and restrictive cells and establishment of latency in mice. J Virol. 1985 Aug;55(2):338–346. doi: 10.1128/jvi.55.2.338-346.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith R. H., Caughman G. B., O'Callaghan D. J. Characterization of the regulatory functions of the equine herpesvirus 1 immediate-early gene product. J Virol. 1992 Feb;66(2):936–945. doi: 10.1128/jvi.66.2.936-945.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith R. H., Holden V. R., O'Callaghan D. J. Nuclear localization and transcriptional activation activities of truncated versions of the immediate-early gene product of equine herpesvirus 1. J Virol. 1995 Jun;69(6):3857–3862. doi: 10.1128/jvi.69.6.3857-3862.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith R. H., Zhao Y., O'Callaghan D. J. The equine herpesvirus 1 (EHV-1) UL3 gene, an ICP27 homolog, is necessary for full activation of gene expression directed by an EHV-1 late promoter. J Virol. 1993 Feb;67(2):1105–1109. doi: 10.1128/jvi.67.2.1105-1109.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith R. H., Zhao Y., O'Callaghan D. J. The equine herpesvirus type 1 immediate-early gene product contains an acidic transcriptional activation domain. Virology. 1994 Aug 1;202(2):760–770. doi: 10.1006/viro.1994.1398. [DOI] [PubMed] [Google Scholar]
- Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
- Telford E. A., Watson M. S., McBride K., Davison A. J. The DNA sequence of equine herpesvirus-1. Virology. 1992 Jul;189(1):304–316. doi: 10.1016/0042-6822(92)90706-u. [DOI] [PubMed] [Google Scholar]
- Tyler J. K., Allen K. E., Everett R. D. Mutation of a single lysine residue severely impairs the DNA recognition and regulatory functions of the VZV gene 62 transactivator protein. Nucleic Acids Res. 1994 Feb 11;22(3):270–278. doi: 10.1093/nar/22.3.270. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vlcek C., Paces V., Schwyzer M. Nucleotide sequence of the pseudorabies virus immediate early gene, encoding a strong transactivator protein. Virus Genes. 1989 Aug;2(4):335–346. doi: 10.1007/BF00684041. [DOI] [PubMed] [Google Scholar]
- Wagner S., Green M. R. HTLV-I Tax protein stimulation of DNA binding of bZIP proteins by enhancing dimerization. Science. 1993 Oct 15;262(5132):395–399. doi: 10.1126/science.8211160. [DOI] [PubMed] [Google Scholar]
- Watson R. J., Clements J. B. A herpes simplex virus type 1 function continuously required for early and late virus RNA synthesis. Nature. 1980 May 29;285(5763):329–330. doi: 10.1038/285329a0. [DOI] [PubMed] [Google Scholar]
- Watson R. J., Preston C. M., Clements J. B. Separation and characterization of herpes simplex virus type 1 immediate-early mRNA's. J Virol. 1979 Jul;31(1):42–52. doi: 10.1128/jvi.31.1.42-52.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whalley J. M., Robertson G. R., Davison A. J. Analysis of the genome of equine herpesvirus type 1: arrangement of cleavage sites for restriction endonucleases EcoRI, BglII and BamHI. J Gen Virol. 1981 Dec;57(Pt 2):307–323. doi: 10.1099/0022-1317-57-2-307. [DOI] [PubMed] [Google Scholar]
- Williams J. S., Andrisani O. M. The hepatitis B virus X protein targets the basic region-leucine zipper domain of CREB. Proc Natl Acad Sci U S A. 1995 Apr 25;92(9):3819–3823. doi: 10.1073/pnas.92.9.3819. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wu C. L., Wilcox K. W. Codons 262 to 490 from the herpes simplex virus ICP4 gene are sufficient to encode a sequence-specific DNA binding protein. Nucleic Acids Res. 1990 Feb 11;18(3):531–538. doi: 10.1093/nar/18.3.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhang G., Leader D. P. The structure of the pseudorabies virus genome at the end of the inverted repeat sequences proximal to the junction with the short unique region. J Gen Virol. 1990 Oct;71(Pt 10):2433–2441. doi: 10.1099/0022-1317-71-10-2433. [DOI] [PubMed] [Google Scholar]
- Zhao Y., Holden V. R., Harty R. N., O'Callaghan D. J. Identification and transcriptional analyses of the UL3 and UL4 genes of equine herpesvirus 1, homologs of the ICP27 and glycoprotein K genes of herpes simplex virus. J Virol. 1992 Sep;66(9):5363–5372. doi: 10.1128/jvi.66.9.5363-5372.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhao Y., Holden V. R., Smith R. H., O'Callaghan D. J. Regulatory function of the equine herpesvirus 1 ICP27 gene product. J Virol. 1995 May;69(5):2786–2793. doi: 10.1128/jvi.69.5.2786-2793.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]