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. 1986 Jul;6(7):2371–2381. doi: 10.1128/mcb.6.7.2371

A mutant herpesvirus protein leads to a block in nuclear localization of other viral proteins.

D M Knipe, J L Smith
PMCID: PMC367790  PMID: 3023931

Abstract

The herpes simplex virus mutants KOS1.1 ts756 and HFEM tsLB2 express temperature-sensitive ICP4 proteins that are not localized properly to the cell nucleus at the nonpermissive temperature. In these infected cells at the nonpermissive temperature, nuclear localization of at least two other viral proteins, ICP0 and ICP8, is impaired. Replacement of the mutated sequences in the ICP4 gene of tsLB2 restored proper nuclear localization of all of the proteins. The ICP0 and ICP8 proteins expressed in cells transfected with their individual genes were localized to the cell nucleus. Therefore, in infected cells, the mutant ICP4 gene product appears to be the primary defect which leads to the block in nuclear localization of the other proteins. One viral protein, ICP27, was not inhibited for nuclear localization in these cells. These data indicate that there are at least two pathways for nuclear localization of HSV proteins, one of which is inhibited by the mutant ICP4 protein. The mutant ICP4 protein may define a probe for one of the pathways of nuclear localization of proteins.

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Selected References

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  1. Ackermann M., Braun D. K., Pereira L., Roizman B. Characterization of herpes simplex virus 1 alpha proteins 0, 4, and 27 with monoclonal antibodies. J Virol. 1984 Oct;52(1):108–118. doi: 10.1128/jvi.52.1.108-118.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ben-Ze'ev A., Abulafia R., Bratosin S. Herpes simplex virus and protein transport are associated with the cytoskeletal framework and the nuclear matrix in infected BSC-1 cells. Virology. 1983 Sep;129(2):501–507. doi: 10.1016/0042-6822(83)90190-3. [DOI] [PubMed] [Google Scholar]
  3. Conley A. J., Knipe D. M., Jones P. C., Roizman B. Molecular genetics of herpes simplex virus. VII. Characterization of a temperature-sensitive mutant produced by in vitro mutagenesis and defective in DNA synthesis and accumulation of gamma polypeptides. J Virol. 1981 Jan;37(1):191–206. doi: 10.1128/jvi.37.1.191-206.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dingwall C., Sharnick S. V., Laskey R. A. A polypeptide domain that specifies migration of nucleoplasmin into the nucleus. Cell. 1982 Sep;30(2):449–458. doi: 10.1016/0092-8674(82)90242-2. [DOI] [PubMed] [Google Scholar]
  5. Dixon R. A., Schaffer P. A. Fine-structure mapping and functional analysis of temperature-sensitive mutants in the gene encoding the herpes simplex virus type 1 immediate early protein VP175. J Virol. 1980 Oct;36(1):189–203. doi: 10.1128/jvi.36.1.189-203.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Everett R. D. Trans activation of transcription by herpes virus products: requirement for two HSV-1 immediate-early polypeptides for maximum activity. EMBO J. 1984 Dec 20;3(13):3135–3141. doi: 10.1002/j.1460-2075.1984.tb02270.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fanning E., Nowak B., Burger C. Detection and characterization of multiple forms of simian virus 40 large T antigen. J Virol. 1981 Jan;37(1):92–102. doi: 10.1128/jvi.37.1.92-102.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gelman I. H., Silverstein S. Identification of immediate early genes from herpes simplex virus that transactivate the virus thymidine kinase gene. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5265–5269. doi: 10.1073/pnas.82.16.5265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ginsberg H. S., Ensinger M. J., Kauffman R. S., Mayer A. J., Lundholm U. Cell transformation: a study of regulation with types 5 and 12 adenovirus temperature-sensitive mutants. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):419–426. doi: 10.1101/sqb.1974.039.01.054. [DOI] [PubMed] [Google Scholar]
  10. Godowski P. J., Knipe D. M. Transcriptional control of herpesvirus gene expression: gene functions required for positive and negative regulation. Proc Natl Acad Sci U S A. 1986 Jan;83(2):256–260. doi: 10.1073/pnas.83.2.256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldstein L., Ko C. Distribution of proteins between nucleus and cytoplasm of Amoeba proteus. J Cell Biol. 1981 Mar;88(3):516–525. doi: 10.1083/jcb.88.3.516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Greenspan D. S., Carroll R. B. Complex of simian virus 40 large tumor antigen and 48,000-dalton host tumor antigen. Proc Natl Acad Sci U S A. 1981 Jan;78(1):105–109. doi: 10.1073/pnas.78.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Halliburton I. W., Randall R. E., Killington R. A., Watson D. H. Some properties of recombinants between type 1 and type 2 herpes simplex viruses. J Gen Virol. 1977 Sep;36(3):471–484. doi: 10.1099/0022-1317-36-3-471. [DOI] [PubMed] [Google Scholar]
  14. Horwitz M. S., Scharff M. D., Maizel J. V., Jr Synthesis and assembly of adenovirus 2. I. Polypeptide synthesis, assembly of capsomeres, and morphogenesis of the virion. Virology. 1969 Dec;39(4):682–694. doi: 10.1016/0042-6822(69)90006-3. [DOI] [PubMed] [Google Scholar]
  15. Hughes R. G., Jr, Munyon W. H. Temperature-sensitive mutants of herpes simplex virus type 1 defective in lysis but not in transformation. J Virol. 1975 Aug;16(2):275–283. doi: 10.1128/jvi.16.2.275-283.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kalderon D., Richardson W. D., Markham A. F., Smith A. E. Sequence requirements for nuclear location of simian virus 40 large-T antigen. Nature. 1984 Sep 6;311(5981):33–38. doi: 10.1038/311033a0. [DOI] [PubMed] [Google Scholar]
  17. Kauffman R. S., Ginsberg H. S. Characterization of a temperature-sensitive, hexon transport mutant of type 5 adenovirus. J Virol. 1976 Aug;19(2):643–658. doi: 10.1128/jvi.19.2.643-658.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Knipe D. M., Ruyechan W. T., Honess R. W., Roizman B. Molecular genetics of herpes simplex virus: the terminal a sequences of the L and S components are obligatorily identical and constitute a part of a structural gene mapping predominantly in the S component. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4534–4538. doi: 10.1073/pnas.76.9.4534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Knipe D. M., Ruyechan W. T., Roizman B., Halliburton I. W. Molecular genetics of herpes simplex virus: demonstration of regions of obligatory and nonobligatory identity within diploid regions of the genome by sequence replacement and insertion. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3896–3900. doi: 10.1073/pnas.75.8.3896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Knipe D. M., Spang A. E. Definition of a series of stages in the association of two herpesviral proteins with the cell nucleus. J Virol. 1982 Jul;43(1):314–324. doi: 10.1128/jvi.43.1.314-324.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lanford R. E., Butel J. S. Inhibition of nuclear migration of wild-type SV40 tumor antigen by a transport-defective mutant of SV40-adenovirus 7 hybrid virus. Virology. 1980 Sep;105(2):303–313. doi: 10.1016/0042-6822(80)90032-x. [DOI] [PubMed] [Google Scholar]
  22. Leibowitz J., Horwitz M. S. Synthesis and assembly of adenovirus polypeptides. III. Reversible inhibition of hexon assembly in adenovirus type 5 temperature-sensitive mutants. Virology. 1975 Jul;66(1):10–24. doi: 10.1016/0042-6822(75)90175-0. [DOI] [PubMed] [Google Scholar]
  23. Metzler D. W., Wilcox K. W. Isolation of herpes simplex virus regulatory protein ICP4 as a homodimeric complex. J Virol. 1985 Aug;55(2):329–337. doi: 10.1128/jvi.55.2.329-337.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Morse L. S., Pereira L., Roizman B., Schaffer P. A. Anatomy of herpes simplex virus (HSV) DNA. X. Mapping of viral genes by analysis of polypeptides and functions specified by HSV-1 X HSV-2 recombinants. J Virol. 1978 May;26(2):389–410. doi: 10.1128/jvi.26.2.389-410.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. O'Hare P., Hayward G. S. Evidence for a direct role for both the 175,000- and 110,000-molecular-weight immediate-early proteins of herpes simplex virus in the transactivation of delayed-early promoters. J Virol. 1985 Mar;53(3):751–760. doi: 10.1128/jvi.53.3.751-760.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Oosterom-Dragon E. A., Ginsberg H. S. Characterization of two temperature-sensitive mutants of type 5 adenovirus with mutations in the 100,000-dalton protein gene. J Virol. 1981 Nov;40(2):491–500. doi: 10.1128/jvi.40.2.491-500.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Powell K. L., Littler E., Purifoy D. J. Nonstructural proteins of herpes simplex virus. II. Major virus-specific DNa-binding protein. J Virol. 1981 Sep;39(3):894–902. doi: 10.1128/jvi.39.3.894-902.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Preston C. M. Control of herpes simplex virus type 1 mRNA synthesis in cells infected with wild-type virus or the temperature-sensitive mutant tsK. J Virol. 1979 Jan;29(1):275–284. doi: 10.1128/jvi.29.1.275-284.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Preston V. G., Davison A. J., Marsden H. S., Timbury M. C., Subak-Sharpe J. H., Wilkie N. M. Recombinants between herpes simplex virus types 1 and 2: analyses of genome structures and expression of immediate early polypeptides. J Virol. 1978 Nov;28(2):499–517. doi: 10.1128/jvi.28.2.499-517.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Prives C., Beck Y., Gidoni D., Oren M., Shure H. DNA binding and sedimentation properties of SV40 T antigens synthesized in vivo and in vitro. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 1):123–130. doi: 10.1101/sqb.1980.044.01.014. [DOI] [PubMed] [Google Scholar]
  31. Quinlan M. P., Chen L. B., Knipe D. M. The intranuclear location of a herpes simplex virus DNA-binding protein is determined by the status of viral DNA replication. Cell. 1984 Apr;36(4):857–868. doi: 10.1016/0092-8674(84)90035-7. [DOI] [PubMed] [Google Scholar]
  32. Quinlan M. P., Knipe D. M. A genetic test for expression of a functional herpes simplex virus DNA-binding protein from a transfected plasmid. J Virol. 1985 May;54(2):619–622. doi: 10.1128/jvi.54.2.619-622.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Quinlan M. P., Knipe D. M. Nuclear localization of herpesvirus proteins: potential role for the cellular framework. Mol Cell Biol. 1983 Mar;3(3):315–324. doi: 10.1128/mcb.3.3.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Quinlan M. P., Knipe D. M. Stimulation of expression of a herpes simplex virus DNA-binding protein by two viral functions. Mol Cell Biol. 1985 May;5(5):957–963. doi: 10.1128/mcb.5.5.957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Russell W. C., Skehel J. J., Williams J. F. Characterization of temperature-sensitive mutants of adenovirus type 5: synthesis of polypeptides in infected cells. J Gen Virol. 1974 Aug;24(2):247–259. doi: 10.1099/0022-1317-24-2-247. [DOI] [PubMed] [Google Scholar]
  36. SMITH K. O. RELATIONSHIP BETWEEN THE ENVELOPE AND THE INFECTIVITY OF HERPES SIMPLEX VIRUS. Proc Soc Exp Biol Med. 1964 Mar;115:814–816. doi: 10.3181/00379727-115-29045. [DOI] [PubMed] [Google Scholar]
  37. Sacks W. R., Greene C. C., Aschman D. P., Schaffer P. A. Herpes simplex virus type 1 ICP27 is an essential regulatory protein. J Virol. 1985 Sep;55(3):796–805. doi: 10.1128/jvi.55.3.796-805.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schaffer P. A., Aron G. M., Biswal N., Benyesh-Melnick M. Temperature-sensitive mutants of herpes simplex virus type 1: isolation, complementation and partial characterization. Virology. 1973 Mar;52(1):57–71. doi: 10.1016/0042-6822(73)90398-x. [DOI] [PubMed] [Google Scholar]
  39. Schaffer P. A., Carter V. C., Timbury M. C. Collaborative complementation study of temperature-sensitive mutants of herpes simplex virus types 1 and 2. J Virol. 1978 Sep;27(3):490–504. doi: 10.1128/jvi.27.3.490-504.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Showalter S. D., Zweig M., Hampar B. Monoclonal antibodies to herpes simplex virus type 1 proteins, including the immediate-early protein ICP 4. Infect Immun. 1981 Dec;34(3):684–692. doi: 10.1128/iai.34.3.684-692.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Velicer L. F., Ginsberg H. S. Synthesis, transport, and morphogenesis of type adenovirus capsid proteins. J Virol. 1970 Mar;5(3):338–352. doi: 10.1128/jvi.5.3.338-352.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. WILDY P., RUSSELL W. C., HORNE R. W. The morphology of herpes virus. Virology. 1960 Oct;12:204–222. doi: 10.1016/0042-6822(60)90195-1. [DOI] [PubMed] [Google Scholar]
  43. Watson R. J., Clements J. B. A herpes simplex virus type 1 function continuously required for early and late virus RNA synthesis. Nature. 1980 May 29;285(5763):329–330. doi: 10.1038/285329a0. [DOI] [PubMed] [Google Scholar]
  44. Willians J. F., Young C. S., Austin P. E. Genetic analysis of human adenovirus type 5 in permissive and nonpermissive cells. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):427–437. doi: 10.1101/sqb.1974.039.01.055. [DOI] [PubMed] [Google Scholar]

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