1887

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Volume 71, Issue 6

Two binding sites for the herpes simplex virus type 1 UL9 protein are required for efficient activity of the ori replication origin Free

Abstract

Two sites within the short region origin of DNA replication (ori) in herpes simplex virus type 1 (HSV-1) which bind the product of the UL9 gene have previously been identified. One of these sites (site I) contains an 11 bp sequence which is also present in ori of varicella-zoster virus, and the other (site II) includes a related element differing in two positions. A third sequence (motif III), which lies close to binding site I, differs from the site I element at only a single position. We have deleted specifically each of these three 11 bp sequences from within functional copies of HSV-1 ori and have examined the effects on origin activity and binding of the UL9 protein. Gel retardation analyses confirmed the important roles of the regions deleted from sites I and II in interacting with the UL9 protein. In transient replication assays, copies of ori lacking the site I or II elements exhibited undetectable or residual (4 to 8 %) activity respectively. The UL9 protein did not bind to motif III even in the absence of site I sequences, although removal of the motif III sequence caused a small reduction in ori activity. A single base change which converted the sequence within binding site I to that of motif III was sufficient to abolish both the interaction of the UL9 gene product at this locus and the replicative ability of ori. Therefore, interaction of the UL9 protein with binding site I is essential for origin activity, but the presence of binding site II is also required for efficient replication.

  • Received:
  • Accepted:
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© Society for General Microbiology 1990
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1990-06-01
2024-04-24
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References

  1. Challberg M. D., Kelly T. J. 1989; Animal virus DNA replication. Annual Review of Biochemistry 58:671–717
     [Google Scholar]
  2. Deb S., Deb S. P. 1989; Analysis of oris sequence of HSV-1: identification of one functional DNA binding domain. Nucleic Acids Research 17:2733–2752
     [Google Scholar]
  3. Deb S., Doelberg M. 1988; A 67-base pair segment from the oris region of herpes simplex virus type 1 encodes origin function. Journal of Virology 62:2516–2519
     [Google Scholar]
  4. 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, U.S.A 85:2959–2963
     [Google Scholar]
  5. Hattori M., Sakaki Y. 1986; Dideoxy sequencing method using denatured plasmid templates. Analytical Biochemistry 152:232–238
     [Google Scholar]
  6. Hubenthal-Voss J., Roizman B. 1988; Properties of two 5′-coterminal RNAs transcribed part way and across the S component origin of DNA synthesis of the herpes simplex virus 1 genome. Proceedings of the National Academy of Sciences, U.S.A. 85:8454–8458
     [Google Scholar]
  7. Hubenthal-Voss J., Starr L., Roizman B. 1987; The herpes simplex virus origins of DNA synthesis in the S component are each contained in a transcribed open reading frame. Journal of Virology 61:3349–3355
     [Google Scholar]
  8. 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]
  9. Kunkel T. A. 1985; Rapid and efficient site-specific mutagenesis without phenotypic selection. Proceedings of the National Academy of Sciences, U.S.A 82:488–492
     [Google Scholar]
  10. Lockshon D., Galloway D. A. 1988; Sequence and structural requirements of a herpes simplex viral DNA replication origin. Molecular and Cellular Biology 8:4018–4027
     [Google Scholar]
  11. Mcgeoch D. J., Dalrymple M. A., Davison A. J., Dolan A., Frame M. C., Mcnab D., Perry L. J., Scott J. E., Taylor P. 1988a; 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]
  12. Mcgeoch D. J., Dalrymple M. A., Dolan A., Mcnab D., Perry L. J., Taylor P., Challberg M. D. 1988b; Structures of herpes simplex virus type 1 genes required for replication of virus DNA. Journal of Virology 62:444–453
     [Google Scholar]
  13. Macpherson I., Stoker M. 1962; Polyoma transformation of hamster cell clones - an investigation of genetic factors affecting cell competence. Virology 16:147–151
     [Google Scholar]
  14. 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]
  15. Mocarski E. S., Roizman B. 1982; Herpes-dependent amplification and inversion of cell-associated viral thymidine kinase gene flanked by viral a sequences and linked to an origin of viral DNA replication. Proceedings of the National Academy of Sciences, U.S.A 79:5626–5630
     [Google Scholar]
  16. 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, U.S.A 85:5414–5418
     [Google Scholar]
  17. Spaete R. R., Frenkel N. 1982; The herpes simplex virus amplicon: a new eucaryotic defective-virus cloning-amplifying vector. Cell 30:295–304
     [Google Scholar]
  18. Stow N. D. 1982; Localization of an origin of DNA replication within the TRS/IRS repeated region of the herpes simplex virus type 1 genome. EMBO Journal 1:863–867
     [Google Scholar]
  19. Stow N. D., Davison A. J. 1986; Identification of a varicella- zoster virus origin of DNA replication and its activation by herpes simplex virus type 1 gene products. Journal of General Virology 67:1613–1623
     [Google Scholar]
  20. Stow N. D., Mcmonagle E. C. 1983; Characterization of the TRS/IRS origin of DNA replication of herpes simplex virus type 1. Virology 130:427–438
     [Google Scholar]
  21. Weir H. M., Calder J. M., Stow N. D. 1989; Binding of the herpes simplex virus type 1 UL9 gene product to an origin of viral DNA replication. Nucleic Acids Research 17:1409–1425
     [Google Scholar]
  22. 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]
  23. Wu C. A., Nelson N. J., Mcgeoch D. J., Challberg M. D. 1988; Identification of herpes simplex virus type 1 genes required for origin-dependent DNA synthesis. Journal of Virology 62:435–443
     [Google Scholar]
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Two binding sites for the herpes simplex virus type 1 UL9 protein are required for efficient activity of the oriS replication origin
J. Gen. Virol. 71, 1379 (1990); https://doi.org/10.1099/0022-1317-71-6-1379
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