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. 1994 Jun;68(6):3841–3849. doi: 10.1128/jvi.68.6.3841-3849.1994

Expression of the varicella-zoster virus origin-binding protein and analysis of its site-specific DNA-binding properties.

D Chen 1, P D Olivo 1
PMCID: PMC236889  PMID: 8189521

Abstract

The varicella-zoster virus (VZV) genome contains homologs to each of the seven herpes simplex virus (HSV) genes that are required for viral DNA synthesis. VZV gene 51 is homologous to HSV UL9, which encodes an origin of DNA replication binding protein (OBP). It was previously shown, by using a protein A fusion protein, that the product of gene 51 is a site-specific DNA-binding protein which binds to sequences within the VZV origin (Stow et al., Virology 177:570-577, 1990). In this report, gene 51 was expressed in an in vitro translation system. Rabbit antiserum raised against the carboxyl-terminal 20 amino acids was used to confirm expression of the full-length gene 51 protein, and site-specific DNA-binding activity was demonstrated in a gel retardation assay. The origin-binding domain was located within a 263-amino-acid region of the carboxyl terminus by using a series of deletion mutants. The affinity of binding of the VZV OBP to the three binding sites in the VZV origin was found to be similar. In addition, as with UL9, a CGC triplet within a 10-bp consensus sequence is critical to the interaction between the OBP and the origin. The HSV and VZV OBPs, therefore, appear to have virtually identical recognition sequences despite only 33% identity and 44% similarity in the primary structure of their site-specific DNA-binding domains.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arbuckle M. I., Stow N. D. A mutational analysis of the DNA-binding domain of the herpes simplex virus type 1 UL9 protein. J Gen Virol. 1993 Jul;74(Pt 7):1349–1355. doi: 10.1099/0022-1317-74-7-1349. [DOI] [PubMed] [Google Scholar]
  2. Challberg M. D. A method for identifying the viral genes required for herpesvirus DNA replication. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9094–9098. doi: 10.1073/pnas.83.23.9094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Challberg M. D., Kelly T. J. Animal virus DNA replication. Annu Rev Biochem. 1989;58:671–717. doi: 10.1146/annurev.bi.58.070189.003323. [DOI] [PubMed] [Google Scholar]
  4. Classification and nomenclature of viruses. Fourth report of the International Committee on Taxonomy of Viruses. Intervirology. 1982;17(1-3):1–199. doi: 10.1159/000149278. [DOI] [PubMed] [Google Scholar]
  5. Davison A. J., Scott J. E. DNA sequence of the major inverted repeat in the varicella-zoster virus genome. J Gen Virol. 1985 Feb;66(Pt 2):207–220. doi: 10.1099/0022-1317-66-2-207. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Deb S., Deb S. P. A 269-amino-acid segment with a pseudo-leucine zipper and a helix-turn-helix motif codes for the sequence-specific DNA-binding domain of herpes simplex virus type 1 origin-binding protein. J Virol. 1991 Jun;65(6):2829–2838. doi: 10.1128/jvi.65.6.2829-2838.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Elias P., Gustafsson C. M., Hammarsten O., Stow N. D. 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. J Biol Chem. 1992 Aug 25;267(24):17424–17429. [PubMed] [Google Scholar]
  9. Elias P., Gustafsson C. M., Hammarsten O. The origin binding protein of herpes simplex virus 1 binds cooperatively to the viral origin of replication oris. J Biol Chem. 1990 Oct 5;265(28):17167–17173. [PubMed] [Google Scholar]
  10. Elias P., O'Donnell M. E., Mocarski E. S., Lehman I. R. A DNA binding protein specific for an origin of replication of herpes simplex virus type 1. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6322–6326. doi: 10.1073/pnas.83.17.6322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hazuda D. J., Perry H. C., McClements W. L. Cooperative interactions between replication origin-bound molecules of herpes simplex virus origin-binding protein are mediated via the amino terminus of the protein. J Biol Chem. 1992 Jul 15;267(20):14309–14315. [PubMed] [Google Scholar]
  12. Hazuda D. J., Perry H. C., Naylor A. M., McClements W. L. Characterization of the herpes simplex virus origin binding protein interaction with OriS. J Biol Chem. 1991 Dec 25;266(36):24621–24626. [PubMed] [Google Scholar]
  13. Martin D. W., Muñoz R. M., Oliver D., Subler M. A., Deb S. Analysis of the DNA-binding domain of the HSV-1 origin-binding protein. Virology. 1994 Jan;198(1):71–80. doi: 10.1006/viro.1994.1009. [DOI] [PubMed] [Google Scholar]
  14. Martinez R., Shao L., Weller S. K. The conserved helicase motifs of the herpes simplex virus type 1 origin-binding protein UL9 are important for function. J Virol. 1992 Nov;66(11):6735–6746. doi: 10.1128/jvi.66.11.6735-6746.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McGeoch D. J., Dalrymple M. A., Dolan A., McNab D., Perry L. J., Taylor P., Challberg M. D. Structures of herpes simplex virus type 1 genes required for replication of virus DNA. J Virol. 1988 Feb;62(2):444–453. doi: 10.1128/jvi.62.2.444-453.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moss B., Elroy-Stein O., Mizukami T., Alexander W. A., Fuerst T. R. Product review. New mammalian expression vectors. Nature. 1990 Nov 1;348(6296):91–92. doi: 10.1038/348091a0. [DOI] [PubMed] [Google Scholar]
  17. Olivo P. D., Nelson N. J., Challberg M. D. Herpes simplex virus DNA replication: the UL9 gene encodes an origin-binding protein. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5414–5418. doi: 10.1073/pnas.85.15.5414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Olivo P. D., Nelson N. J., Challberg M. D. Herpes simplex virus type 1 gene products required for DNA replication: identification and overexpression. J Virol. 1989 Jan;63(1):196–204. doi: 10.1128/jvi.63.1.196-204.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Spaete R. R., Frenkel N. The herpes simplex virus amplicon: a new eucaryotic defective-virus cloning-amplifying vector. Cell. 1982 Aug;30(1):295–304. doi: 10.1016/0092-8674(82)90035-6. [DOI] [PubMed] [Google Scholar]
  20. Stillman B. Initiation of eukaryotic DNA replication in vitro. Annu Rev Cell Biol. 1989;5:197–245. doi: 10.1146/annurev.cb.05.110189.001213. [DOI] [PubMed] [Google Scholar]
  21. Stow N. D., Davison A. J. Identification of a varicella-zoster virus origin of DNA replication and its activation by herpes simplex virus type 1 gene products. J Gen Virol. 1986 Aug;67(Pt 8):1613–1623. doi: 10.1099/0022-1317-67-8-1613. [DOI] [PubMed] [Google Scholar]
  22. Stow N. D. Localization of an origin of DNA replication within the TRS/IRS repeated region of the herpes simplex virus type 1 genome. EMBO J. 1982;1(7):863–867. doi: 10.1002/j.1460-2075.1982.tb01261.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Stow N. D., Weir H. M., Stow E. C. Analysis of the binding sites for the varicella-zoster virus gene 51 product within the viral origin of DNA replication. Virology. 1990 Aug;177(2):570–577. doi: 10.1016/0042-6822(90)90522-s. [DOI] [PubMed] [Google Scholar]
  24. Vlazny D. A., Frenkel N. Replication of herpes simplex virus DNA: localization of replication recognition signals within defective virus genomes. Proc Natl Acad Sci U S A. 1981 Feb;78(2):742–746. doi: 10.1073/pnas.78.2.742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Weller S. K., Spadaro A., Schaffer J. E., Murray A. W., Maxam A. M., Schaffer P. A. Cloning, sequencing, and functional analysis of oriL, a herpes simplex virus type 1 origin of DNA synthesis. Mol Cell Biol. 1985 May;5(5):930–942. doi: 10.1128/mcb.5.5.930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wu C. A., Nelson N. J., McGeoch D. J., Challberg M. D. Identification of herpes simplex virus type 1 genes required for origin-dependent DNA synthesis. J Virol. 1988 Feb;62(2):435–443. doi: 10.1128/jvi.62.2.435-443.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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