1887

Abstract

Herpes simplex virus type 1 expresses a heterotrimeric helicase–primase, the subunits of which are encoded by the viral , and genes. The interactions of the UL52 protein with the UL8 and UL5 proteins were analysed by using the yeast two-hybrid system. The UL52–UL5 interaction gave a specific but weak signal in the two-hybrid system. In contrast, the UL52–UL8 interaction gave a strong signal in the two-hybrid system. Deletion analysis showed that a 548 amino acid fragment of UL52 (amino acids 366–914) retains the ability to interact with UL8 and that the N-terminal 349 amino acids are dispensable for the interaction. A fragment library screen and co-immunoprecipitation experiments confirmed the deletion analysis results.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-80-9-2411
1999-09-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/80/9/0802411a.html?itemId=/content/journal/jgv/10.1099/0022-1317-80-9-2411&mimeType=html&fmt=ahah

References

  1. Barnard E. C., Brown G., Stow N. D. 1997; Deletion mutants of the herpes simplex virus type 1 UL8 protein: effect on DNA synthesis and ability to interact with and influence the intracellular localization of the UL5 and UL52 proteins. Virology 237:97–106
    [Google Scholar]
  2. Bartel P. L., Chien C.-T., Sternglanz R., Fields S. 1993a; Using the two-hybrid system to detect protein–protein interactions. Cellular Interactions in Development: A Practical Approach153–179 Oxford: Oxford University Press;
    [Google Scholar]
  3. Bartel P. L., Chien C.-T., Sternglanz R., Fields S. 1993b; Elimination of false positives that arise in using the two-hybrid system. Biotechniques 14:920–924
    [Google Scholar]
  4. Breeden L., Nasmyth K. 1985; Regulation of the yeast HO gene. Cold Spring Harbor Symposia on Quantitative Biology 50:643–650
    [Google Scholar]
  5. Calder J. M., Stow N. D. 1990; Herpes simplex virus helicase–primase: the UL8 protein is not required for DNA-dependent ATPase and DNA helicase activities. Nucleic Acids Research 18:3573–3578
    [Google Scholar]
  6. Calder J. M., Stow E. C., Stow N. D. 1992; On the cellular localization of the components of the herpes simplex virus type 1 helicase–primase complex and the viral origin-binding protein. Journal of General Virology 73:531–538
    [Google Scholar]
  7. Carmichael E. P., Weller S. K. 1989; Herpes simplex virus type 1 DNA synthesis requires the product of the UL8 gene: isolation and characterization of an ICP6:: lacZ insertion mutation. Journal of Virology 63:591–599
    [Google Scholar]
  8. Challberg M. D. 1986; A method for identifying the viral genes required for herpesvirus DNA replication. Proceedings of the National Academy of Sciences USA 83:9094–9098
    [Google Scholar]
  9. Challberg M. D., Kelly T. J. 1989; Animal virus DNA replication. Annual Review of Biochemistry 58:671–717
    [Google Scholar]
  10. Chien C.-T., Bartel P. L., Sternglanz R., Fields S. 1991; The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proceedings of the National Academy of Sciences USA 889578–9582
    [Google Scholar]
  11. Crute J. J., Lehman I. R. 1991; Herpes simplex virus-1 helicase–primase. Physical and catalytic properties. Journal of Biological Chemistry 266:4484–4488
    [Google Scholar]
  12. Crute J. J., Tsurumi T., Zhu L. A., Weller S. K., Olivo P. D., Challberg M. D., Mocarski E. S., Lehman I. R. 1989; Herpes simplex virus 1 helicase–primase: a complex of three herpes-encoded gene products. Proceedings of the National Academy of Sciences USA 86:2186–2189
    [Google Scholar]
  13. Dodson M. S., Lehman I. R. 1991; Association of DNA helicase and primase activities with a subassembly of the herpes simplex virus 1 helicase–primase composed of the UL5 and UL52 gene products. Proceedings of the National Academy of Sciences USA 88:1105–1109
    [Google Scholar]
  14. Dodson M. S., Crute J. J., Bruckner R. C., Lehman I. R. 1989; Overexpression and assembly of the herpes simplex virus type 1 helicase–primase in insect cells. Journal of Biological Chemistry 264:20835–20838
    [Google Scholar]
  15. Dracheva S., Koonin E. V., Crute J. J. 1995; Identification of the primase active site of the herpes simplex virus type 1 helicase–primase. Journal of Biological Chemistry 270:14148–14153
    [Google Scholar]
  16. Falkenberg M., Bushnell D. A., Elias P., Lehman I. R. 1997; The UL8 subunit of the heterotrimeric herpes simplex virus type 1 helicase–primase is required for the unwinding of single strand DNA-binding protein (ICP8)-coated DNA substrates. Journal of Biological Chemistry 272:22766–22770
    [Google Scholar]
  17. Feilotter H. E., Hannon G. J., Ruddell C. J., Beach D. 1994; Construction of an improved host strain for two hybrid screening. Nucleic Acids Research 22:1502–1503
    [Google Scholar]
  18. Fields S., Song O. 1989; A novel genetic system to detect protein–protein interactions. Nature 340:245–246
    [Google Scholar]
  19. Gorbalenya A. E., Koonin E. 1993; Helicases: amino acid sequence comparisons and structure–function relationships. Current Opinion in Structural Biology 3:419–429
    [Google Scholar]
  20. Graves-Woodward K. L., Weller S. K. 1996; Replacement of gly815 in helicase motif V alters the single-stranded DNA-dependent ATPase activity of the herpes simplex virus type 1 helicase–primase. Journal of Biological Chemistry 271:13629–13635
    [Google Scholar]
  21. Graves-Woodward K. L., Gottlieb J., Challberg M. D., Weller S. K. 1997; Biochemical analyses of mutations in the HSV-1 helicase–primase that alter ATP hydrolysis, DNA unwinding, and coupling between hydrolysis and unwinding. Journal of Biological Chemistry 272:4623–4630
    [Google Scholar]
  22. Klinedinst D. K., Challberg M. D. 1994; Helicase–primase complex of herpes simplex type 1: a mutation in the UL52 subunit abolishes primase activity. Journal of Virology 68:3693–3701
    [Google Scholar]
  23. 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]
  24. McLean G. W., Abbotts A. P., Parry M. E., Marsden H. S., Stow N. D. 1994; The herpes simplex virus type 1 origin-binding protein interacts specifically with the viral UL8 protein. Journal of General Virology 75:2699–2706
    [Google Scholar]
  25. Marsden H. S., Cross A. M., Francis G. J., Patel A. H., MacEachran K., Murphy M., McVey G., Haydon D., Abbotts A., Stow N. D. 1996; The herpes simplex virus type 1 UL8 protein influences the intracellular localization of the UL52 but not the ICP8 or POL replication proteins in virus-infected cells. Journal of General Virology 77:2241–2249
    [Google Scholar]
  26. Marsden H. S., McLean G. W., Barnard E. C., Francis G. J., MacEachran K., Murphy M., McVey G., Cross A., Abbotts A. P., Stow N. D. 1997; The catalytic subunit of the DNA polymerase of herpes simplex virus type 1 interacts specifically with the C terminus of the UL8 component of the viral helicase–primase complex. Journal of Virology 71:6390–6397
    [Google Scholar]
  27. Parry M. E., Stow N. D., Marsden H. S. 1993; Purification and properties of the herpes simplex virus type 1 UL8 protein. Journal of General Virology 74:607–612
    [Google Scholar]
  28. Sherman G., Gottlieb J., Challberg M. D. 1992; The UL8 subunit of the herpes simplex virus helicase–primase complex is required for efficient primer utilization. Journal of Virology 66:4884–4892
    [Google Scholar]
  29. Summers M. D., Smith G. E. 1987 A Manual of Methods for Baculovirus Vectors and Insect Cell Procedures College Station, TX: Texas A&M University;
    [Google Scholar]
  30. Tenney D. J., Hurlburt W. W., Micheletti P. A., Bifano M., Hamatake R. K. 1994; The UL8 component of the herpes simplex virus helicase–primase complex stimulates primer synthesis by a subassembly of the UL5 and UL52 components. Journal of Biological Chemistry 269:5030–5035
    [Google Scholar]
  31. Zhu L. A., Weller S. K. 1992; The UL5 gene of herpes simplex virus type 1: isolation of a lacZ insertion mutant and association of the UL5 gene product with other members of the helicase–primase complex. Journal of Virology 66:458–468
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-80-9-2411
Loading
/content/journal/jgv/10.1099/0022-1317-80-9-2411
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error