1887

Abstract

The UL15 and UL28 proteins of herpes simplex virus type 1 are both required for the packaging of replicated viral DNA into the viral capsid. We have expressed UL28 and a functional epitope-tagged form of UL15 in mammalian and insect cells. Immunoprecipitation experiments confirmed that the two proteins can interact. In agreement with previous results, UL15, when expressed alone, entered the nucleus but UL28 remained cytoplasmic. When co-expressed the two proteins co-localized in the nucleus. Six UL28 deletion mutants were constructed and similarly analysed. The results obtained by immunoprecipitation and immunofluorescence were consistent and demonstrate that at least two separate regions of the UL28 polypeptide chain have the ability to interact with UL15. Surprisingly, three of the mutants prevented the UL15 protein from localizing to the cell nucleus, and these were not functional in a transient DNA packaging assay. Of the three UL28 mutant proteins that entered the nucleus with UL15, one containing an internal deletion of 13 amino acids was able to complement a UL28 null mutant in both DNA packaging and virus yield assays, demonstrating that this region of the protein is not essential for function. In addition to interacting with the UL28 protein we also demonstrated that UL15 molecules can interact with each other, and that sequences within the second exon contribute to this interaction.

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2000-12-01
2019-10-20
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References

  1. Addison, C., Rixon, F. J., Palfreyman, J. W., O’Hara, M. & Preston, V. G. ( 1984; ). Characterisation of a herpes simplex virus type 1 mutant which has a temperature-sensitive defect in penetration of cells and assembly of capsids. Virology 138, 246-259.[CrossRef]
    [Google Scholar]
  2. Addison, C., Rixon, F. J. & Preston, V. G. ( 1990; ). Herpes simplex virus type 1 UL28 gene product is important for the formation of mature capsids. Journal of General Virology 71, 2377-2384.[CrossRef]
    [Google Scholar]
  3. Ali, M. A., Forghani, B. & Cantin, E. M. ( 1996; ). Characterization of an essential HSV-1 protein encoded by the UL25 gene reported to be involved in virus penetration and capsid assembly. Virology 216, 278-283.[CrossRef]
    [Google Scholar]
  4. Baines, J. D., Cunningham, C., Nalwanga, D. & Davison, A. ( 1997; ). The UL15 gene of herpes simplex virus type 1 contains within its second exon a novel open reading frame that is translated in frame with the UL15 gene product. Journal of Virology 71, 2666-2673.
    [Google Scholar]
  5. Bishop, D. H. L. ( 1992; ). Baculovirus expression vectors. Seminars in Virology 3, 253-264.
    [Google Scholar]
  6. Bogner, E., Radsak, K. & Stinski, M. F. ( 1998; ). The gene product of human cytomegalovirus open reading frame UL56 binds the pac motif and has specific nuclease activity. Journal of Virology 72, 2259-2264.
    [Google Scholar]
  7. Catalano, C. E. ( 2000; ). The terminase enzyme from bacteriophage lambda: a DNA-packaging machine. Cellular and Molecular Life Sciences 57, 128-148.[CrossRef]
    [Google Scholar]
  8. Catalano, C. E., Cue, D. & Feiss, M. ( 1995; ). Virus DNA packaging: the strategy used by phage lambda. Molecular Microbiology 16, 1075-1086.[CrossRef]
    [Google Scholar]
  9. Cavalcoli, J. D., Baghian, A., Homa, F. L. & Kousoulas, K. G. ( 1993; ). Resolution of genotypic and phenotypic properties of herpes simplex virus type 1 temperature-sensitive mutant (KOS) tsZ47: evidence for allelic complementation in the UL28 gene. Virology 197, 23-34.[CrossRef]
    [Google Scholar]
  10. Davison, A. J. ( 1992; ). Channel catfish virus: a new type of herpesvirus. Virology 186, 9-14.[CrossRef]
    [Google Scholar]
  11. Dolan, A., Arbuckle, M. & McGeoch, D. J. ( 1991; ). Sequence analysis of the splice junction in the transcript of herpes simplex virus type 1 UL15. Virus Research 20, 97-104.[CrossRef]
    [Google Scholar]
  12. Fujisawa, H. & Morita, M. ( 1997; ). Phage DNA packaging. Genes to Cells 2, 537-545.
    [Google Scholar]
  13. Goshima, F., Watanabe, D., Takakuwa, H., Wada, K., Daikoku, T., Yamada, M. & Nishiyama, Y. ( 2000; ). Herpes simplex virus UL17 protein is associated with B capsids and colocalizes with ICP35 and VP5 in infected cells. Archives of Virology 145, 417-426.[CrossRef]
    [Google Scholar]
  14. Homa, F. L. & Brown, J. C. ( 1997; ). Capsid assembly and DNA packaging in herpes simplex virus. Reviews in Medical Virology 7, 107-122.[CrossRef]
    [Google Scholar]
  15. Kitts, P. A., Ayres, M. D. & Possee, R. D. ( 1990; ). Linearization of baculovirus DNA enhances the recovery of recombinant virus expression vectors. Nucleic Acids Research 18, 5667-5672.[CrossRef]
    [Google Scholar]
  16. Koslowski, K. M., Shaver, P. R., Wang, X.-Y., Tenney, D. J. & Pederson, N. E. ( 1997; ). The pseudorabies virus UL28 protein enters the nucleus after coexpression with the herpes simplex virus UL15 protein. Journal of Virology 71, 9118-9123.
    [Google Scholar]
  17. Koslowski, K. M., Shaver, P. R., Casey, J. T.II, Wilson, T., Yamanaka, G. Y., Sheaffer, A. K., Tenney, D. J. & Pederson, N. E. ( 1999; ). Physical and functional interactions between the herpes simplex virus UL15 and UL28 DNA cleavage and packaging proteins. Journal of Virology 73, 1704-1707.
    [Google Scholar]
  18. Krosky, P. M., Underwood, M. R., Turk, S. R., Feng, K. W.-H., Jain, R. K., Ptak, R. G., Westerman, A. C., Biron, K. K., Townsend, L. B. & Drach, J. C. ( 1998; ). Resistance of human cytomegalovirus to benzimidazole ribonucleosides maps to two open reading frames: UL89 and UL56. Journal of Virology 72, 4721-4728.
    [Google Scholar]
  19. Lai, J.-S. & Herr, W. ( 1992; ). Ethidium bromide provides a simple test for identifying genuine DNA-independent protein associations. Proceedings of the National Academy of Sciences, USA 89, 6958-6962.[CrossRef]
    [Google Scholar]
  20. Lamberti, C. & Weller, S. K. ( 1998; ). The herpes simplex virus type 1 cleavage/packaging protein, UL32, is involved in efficient localization of capsids to replication compartments. Journal of Virology 72, 2463-2473.
    [Google Scholar]
  21. Livingstone, C. & Jones, I. ( 1989; ). Baculovirus expression vectors with single strand capability. Nucleic Acids Research 17, 2366.[CrossRef]
    [Google Scholar]
  22. 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.[CrossRef]
    [Google Scholar]
  23. 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.[CrossRef]
    [Google Scholar]
  24. McNab, A. R., Desai, P., Person, S., Roof, L. L., Thomsen, D. R., Newcomb, W. W., Brown, J. C. & Homa, F. L. ( 1998; ). The product of the herpes simplex virus type 1 UL25 gene is required for encapsidation but not for cleavage of replicated viral DNA. Journal of Virology 72, 1060-1070.
    [Google Scholar]
  25. Malik, A. K., Shao, L., Shanley, J. D. & Weller, S. K. ( 1996; ). Intracellular localization of the herpes simplex virus type 1 origin binding protein, UL9. Virology 224, 380-389.[CrossRef]
    [Google Scholar]
  26. Nasseri, M. & Mocarski, E. S. ( 1988; ). The cleavage recognition signal is contained within sequences surrounding an a–a junction in herpes simplex virus DNA. Virology 167, 25-30.[CrossRef]
    [Google Scholar]
  27. Patel, A. H. & Maclean, J. B. ( 1995; ). The product of the UL6 gene of herpes simplex virus type 1 is associated with virus capsids. Virology 206, 465-478.[CrossRef]
    [Google Scholar]
  28. Poon, A. P. W. & Roizman, B. ( 1993; ). Characterization of a temperature-sensitive mutant of the UL15 open reading frame of herpes simplex virus 1. Journal of Virology 67, 4497-4503.
    [Google Scholar]
  29. Reynolds, A. E., Fan, Y. & Baines, J. D. ( 2000; ). Characterization of the UL33 gene product of herpes simplex virus 1. Virology 266, 310-318.[CrossRef]
    [Google Scholar]
  30. Rixon, F. J. ( 1993; ). Structure and assembly of herpesviruses. Seminars in Virology 4, 135-144.[CrossRef]
    [Google Scholar]
  31. Rosenthal, K. S., Leuther, M. D. & Baines, B. G. ( 1984; ). Herpes simplex virus binding and entry modulate cell surface protein mobility. Journal of Virology 49, 980-983.
    [Google Scholar]
  32. Salmon, B. & Baines, J. D. ( 1998; ). Herpes simplex virus DNA cleavage and packaging: association of multiple forms of UL15-encoded proteins with B capsids requires at least the UL6, UL17 and UL28 genes. Journal of Virology 72, 3045-3050.
    [Google Scholar]
  33. Salmon, B., Cunningham, C., Davison, A. J., Harris, W. J. & Baines, J. D. ( 1998; ). The herpes simplex virus type 1 UL17 gene encodes virion tegument proteins that are required for cleavage and packaging of viral DNA. Journal of Virology 72, 3779-3788.
    [Google Scholar]
  34. Stow, N. D. ( 1998; ). Transient assays for HSV origin and replication protein function. In Herpes Simplex Virus Protocols, pp. 215-226. Edited by S. M. Brown & A. R. MacLean. Totowa, NJ:Humana Press.
  35. 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.[CrossRef]
    [Google Scholar]
  36. Stow, N. D. & Wilkie, N. M. ( 1976; ). An improved technique for obtaining enhanced infectivity with herpes simplex virus type 1 DNA. Journal of General Virology 33, 447-458.[CrossRef]
    [Google Scholar]
  37. Stow, N. D., McMonagle, E. C. & Davison, A. J. ( 1983; ). Fragments from both termini of the herpes simplex virus type 1 genome contain signals required for the encapsidation of viral DNA. Nucleic Acids Research 11, 8205-8220.[CrossRef]
    [Google Scholar]
  38. 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.[CrossRef]
    [Google Scholar]
  39. Stow, N. D., Brown, G., Cross, A. M. & Abbotts, A. P. ( 1998; ). Identification of residues within the herpes simplex virus type 1 origin-binding protein that contribute to sequence-specific DNA binding. Virology 240, 183-192.[CrossRef]
    [Google Scholar]
  40. Taus, N. S. & Baines, J. D. ( 1998; ). Herpes simplex virus DNA cleavage/packaging: the UL28 gene encodes a minor component of B capsids. Virology 252, 443-449.[CrossRef]
    [Google Scholar]
  41. Tengelsen, L. A., Pederson, N. E., Shaver, P. R., Wathen, M. W. & Homa, F. L. ( 1993; ). Herpes simplex virus type 1 DNA cleavage and encapsidation require the product of the UL28 gene: isolation and characterization of two UL28 deletion mutants. Journal of Virology 67, 3470-3480.
    [Google Scholar]
  42. Towbin, H., Staehelin, T. & Gordon, J. ( 1979; ). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, USA 76, 4350-4354.[CrossRef]
    [Google Scholar]
  43. Underwood, M. R., Harvey, R. J., Stanat, S. C., Hemphill, M. L., Miller, T., Drach, J. C., Townsend, L. B. & Biron, K. K. ( 1998; ). Inhibition of human cytomegalovirus DNA maturation by a benzimidazole ribonucleoside is mediated through the UL89 gene product. Journal of Virology 72, 717-725.
    [Google Scholar]
  44. Yu, D. & Weller, S. K. ( 1998a; ). Herpes simplex virus type 1 cleavage and packaging proteins UL15 and UL28 are associated with B but not C capsids during packaging. Journal of Virology 72, 7428-7439.
    [Google Scholar]
  45. Yu, D. & Weller, S. K. ( 1998b; ). Genetic analysis of the UL15 gene locus for the putative terminase of herpes simplex virus type 1. Virology 243, 32-44.[CrossRef]
    [Google Scholar]
  46. Yu, D., Sheaffer, A. K., Tenney, D. J. & Weller, S. K. ( 1997; ). Characterization of ICP6::lacZ insertion mutants of the UL15 gene of herpes simplex virus type 1 reveals the translation of two proteins. Journal of Virology 71, 2656-2665.
    [Google Scholar]
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