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Volume 77, Issue 5

Characterization of a binding site for the herpes simplex virus type 1 UL9 origin-binding protein within the UL9 gene Free

Abstract

Gene UL9 of herpes simplex virus type 1 (HSV-1) encodes a sequence-specific origin-binding protein (OBP) that plays a direct and essential role in viral DNA synthesis. A search of the complete HSV-1 genomic sequence for possible OBP binding sites lying outside the known origins of replication revealed the presence of a very close match to the OBP recognition sequence within the UL9 coding region. The ability of OBP to bind to this site (referred to as the ‘UL9 box’) was confirmed by DNase I footprinting and gel retardation assays, and filter binding experiments demonstrated that the affinity of OBP for the UL9 box was of the same order as for its high affinity sites within the three replication origins. To investigate whether binding of OBP to the UL9 box played a role during viral replication we constructed a mutant virus in which the sequence was altered in such a way as to preserve the encoded amino acid sequence whilst abolishing the ability of OBP to bind. Growth of the virus was indistinguishable from wild-type and no alterations were observed in the accumulation of transcripts from the UL9 region of the genome. In addition, a DNA fragment containing the UL9 box sequence did not exhibit origin activity in a transient assay for viral DNA synthesis. We therefore conclude that binding of OBP to the UL9 box is not essential for virus growth and that expression of the UL9 gene is unlikely to be autoregulated through this site.

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© Journal of General Virology 1996
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1996-05-01
2024-04-19
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References

  1. Baradaran K., Dabrowski C. E., Schaffer P. A. 1994; Transcriptional analysis of the region of the herpes simplex virus type 1 genome containing the UL8, UL9 and UL10 genes and identification of a novel delayed-early gene product, OBPC. Journal of Virology 68:4251–4261
     [Google Scholar]
  2. Blumel J., Matz B. 1995; Thermosensitive UL9 gene function is required for early stages of herpes simplex virus type 1 DNA synthesis. Journal of General Virology 76:3119–3124
     [Google Scholar]
  3. Brown S. M., Ritchie D. A., Subak-Sharpe J. H. 1973; Genetic studies with herpes simplex virus type 1. The isolation of temperature-sensitive mutants, their arrangement into complementation groups and recombination analysis leading to a linkage map. Journal of General Virology 18:329–346
     [Google Scholar]
  4. Challberg M. D. 1991; Herpes simplex virus DNA replication. Seminars in Virology 2:247–256
     [Google Scholar]
  5. Chomczynski P., Sacchi N. 1987; Single step method of RNA isolation by guanidinium thiocyanate-phenol-chloroform extraction. Annals of Biochemistry 162:156–159
     [Google Scholar]
  6. Dabrowski C. E., Schaffer P. A. 1991; Herpes simplex virus type-1 origin-specific binding protein: oris-binding properties and effects of cellular proteins. Journal of Virology 65:3140–3150
     [Google Scholar]
  7. Dabrowski C. E., Carmillo P. J., Schaffer P. A. 1994; Cellular protein interactions with herpes simplex virus type 1 oris . Molecular and Cellular Biology 14:2545–2555
     [Google Scholar]
  8. Davison A. J., Scott J. E. 1986; The complete DNA sequence of varicella-zoster virus. Journal of General Virology 67:1759–1816
     [Google Scholar]
  9. Elias P., Lehman I. R. 1988; Interaction of origin binding protein with an origin of replication of herpes simplex virus 1. Proceedings of the National Academy of Sciences, USA 85:2959–2963
     [Google Scholar]
  10. Elias P., Gustafsson C. M., Hammarsten O. 1990; The origin binding protein of herpes simplex virus 1 binds cooperatively to the viral origin of replication oris . Journal of Biological Chemistry 265:17167–17173
     [Google Scholar]
  11. Elias P., Gustafsson C. M., Hammarsten O., Stow N. D. 1992; Structural elements required for the cooperative binding of the herpes simplex virus origin binding protein to oris reside in the N- terminal part of the protein. Journal of Biological Chemistry 267:17424–17429
     [Google Scholar]
  12. Hazuda D. J., Perry H. C., Naylor A. M., McClements W. L. 1991; Characterization of the herpes simplex virus origin binding protein interaction with oriS. Journal of Biological Chemistry 266:24621–24626
     [Google Scholar]
  13. Hernandez T. R., Dutch R. E., Lehman I. R., Gustafsson C., Elias P. 1991; Mutations in a herpes simplex virus type 1 origin that inhibit interaction with origin-binding protein also inhibit DNA replication. Journal of Virology 65:1649–1652
     [Google Scholar]
  14. Koff A., Tegtmeyer P. 1988; Characterization of major recognition sequences for a herpes simplex virus type 1 originbinding protein. Journal of Virology 62:4096–4103
     [Google Scholar]
  15. McGeoch D. J., Dalrymple M. A., Davison A. J., Dolan A., Frame M. C., McNab D., Perry L. J., Scott J. E., Taylor P. 1988; The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. Journal of General Virology 69:1531–1574
     [Google Scholar]
  16. Malik A. K., Martinez R., Muncy L., Carmichael E. P., Weller S. K. 1992; Genetic analysis of the herpes simplex virus type 1 UL9 gene: isolation of a lacZ insertion mutant and expression in eukaryotic cells. Virology 190:702–715
     [Google Scholar]
  17. Marsden H. S., Crombie I. K., Subak-Sharpe J. H. 1976; Control of protein synthesis in herpesvirus-infected cells: analysis of polypeptides induced by wild type and sixteen temperature-sensitive mutants of HSV strain 17. Journal of General Virology 31:347–372
     [Google Scholar]
  18. Martin D. W., Deb S. 1994; Cloning and expression of an equine herpesvirus-1 origin-binding protein. Journal of Virology 68:3674–3681
     [Google Scholar]
  19. Martin D. W., Deb S. P., Klauer J. S., Deb S. 1991; Analysis of the herpes simplex virus type 1 oris sequence: mapping of functional domains. Journal of Virology 65:4359–4369
     [Google Scholar]
  20. Mead D. A., Szczesna-Skorupa E., Kemper B. 1986; Single stranded DNA ‘blue’ T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Engineering 1:67–74
     [Google Scholar]
  21. Olivo P. D., Nelson N. J., Challberg M. D. 1988; Herpes simplex virus DNA replication: the UL9 gene encodes an originbinding protein. Proceedings of the National Academy of Sciences, USA 85:5414–5418
     [Google Scholar]
  22. Stow N. D., McMonagle E. C. 1983; Characterization of the TRa/IRs origin of DNA replication of herpes simplex virus type 1. Virology 130:427–438
     [Google Scholar]
  23. Stow N. D., Weir H. M., Stow E. C. 1990; Analysis of the binding sites for the varicella-zoster virus gene 51 product within the viral origin of DNA replication. Virology 177:570–577
     [Google Scholar]
  24. Stow N. D., Subak-Sharpe J. H., Wilkie N. M. 1978; Physical mapping of herpes simplex virus type 1 mutations by marker rescue. Journal of Virology 28:182–192
     [Google Scholar]
  25. Stow N. D., Hammarsten O., Arbuckle M. I., Elias P. 1993; Inhibition of herpes simplex virus type 1 DNA replication by mutant forms of the origin-binding protein. Virology 196:413–418
     [Google Scholar]
  26. Telford E. A. R., Watson M. S., McBride K., Davison A. J. 1992; The DNA sequence of equine herpesvirus-1. Virology 189:304–316
     [Google Scholar]
  27. Weir H. M., Stow N. D. 1990; Two binding sites for the herpes simplex virus type 1 UL9 gene product are required for efficient activity of the oris replication origin. Journal of General Virology 71:1379–1385
     [Google Scholar]
  28. Weller S. K., Spadaro A., Schaffer J. E., Murray A. W., Maxam A. M., Schaffer P. A. 1985; Cloning, sequencing, and functional analysis of oriL, a herpes simplex virus type 1 origin of DNA synthesis. Molecular and Cellular Biology 5:930–942
     [Google Scholar]
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Characterization of a binding site for the herpes simplex virus type 1 UL9 origin-binding protein within the UL9 gene
J. Gen. Virol. 77, 969 (1996); https://doi.org/10.1099/0022-1317-77-5-969
/content/journal/jgv/10.1099/0022-1317-77-5-969
/content/journal/jgv/10.1099/0022-1317-77-5-969
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