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. 1993 Dec;67(12):7149–7160. doi: 10.1128/jvi.67.12.7149-7160.1993

Isolation of a herpes simplex virus type 1 mutant with a deletion in the virion host shutoff gene and identification of multiple forms of the vhs (UL41) polypeptide.

G S Read 1, B M Karr 1, K Knight 1
PMCID: PMC238177  PMID: 8230437

Abstract

The virion host shutoff (vhs) gene (UL41) of herpes simplex virus type 1 (HSV-1) encodes a virion component that induces degradation of host mRNAs and the shutoff of most host protein synthesis. Subsequently, the vhs protein accelerates the turnover of all kinetic classes of viral mRNA. To identify the vhs (UL41) polypeptide within infected cells and virions, antisera raised against a UL41-lacZ fusion protein were used to characterize the polypeptides encoded by wild-type HSV-1 and two mutants: vhs1, a previously characterized mutant that lacks detectable virion host shutoff activity, and vhs-delta Sma, a newly constructed mutant containing a deletion of 196 codons from UL41. Two forms of the vhs (UL41) polypeptide were identified in cells infected with the wild-type virus or vhs1. Wild-type HSV-1 produced a major 58-kDa polypeptide, as well as a less abundant 59.5-kDa form of the protein, while vhs1 produced 57- and 59-kDa polypeptides that were approximately equally abundant. Although for either virus, both forms of the protein were phosphorylated, they differed in the extent of phosphorylation. While both vhs polypeptides were found in infected cells, only the faster migrating, less phosphorylated form was incorporated into virions. vhs-delta Sma encoded a smaller, 31-kDa polypeptide which, although present in infected cells, was not incorporated into virions. The results identify multiple forms of the vhs (UL41) polypeptide and suggest that posttranslational processing affects its packaging into virions, as well as its ability to induce mRNA degradation.

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Selected References

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

  1. Anderson K. P., Frink R. J., Devi G. B., Gaylord B. H., Costa R. H., Wagner E. K. Detailed characterization of the mRNA mapping in the HindIII fragment K region of the herpes simplex virus type 1 genome. J Virol. 1981 Mar;37(3):1011–1027. doi: 10.1128/jvi.37.3.1011-1027.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Atwater J. A., Wisdom R., Verma I. M. Regulated mRNA stability. Annu Rev Genet. 1990;24:519–541. doi: 10.1146/annurev.ge.24.120190.002511. [DOI] [PubMed] [Google Scholar]
  3. Bell S., Cranage M., Borysiewicz L., Minson T. Induction of immunoglobulin G Fc receptors by recombinant vaccinia viruses expressing glycoproteins E and I of herpes simplex virus type 1. J Virol. 1990 May;64(5):2181–2186. doi: 10.1128/jvi.64.5.2181-2186.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bernstein P., Ross J. Poly(A), poly(A) binding protein and the regulation of mRNA stability. Trends Biochem Sci. 1989 Sep;14(9):373–377. doi: 10.1016/0968-0004(89)90011-x. [DOI] [PubMed] [Google Scholar]
  5. Brawerman G. mRNA decay: finding the right targets. Cell. 1989 Apr 7;57(1):9–10. doi: 10.1016/0092-8674(89)90166-9. [DOI] [PubMed] [Google Scholar]
  6. Buchkovich K., Duffy L. A., Harlow E. The retinoblastoma protein is phosphorylated during specific phases of the cell cycle. Cell. 1989 Sep 22;58(6):1097–1105. doi: 10.1016/0092-8674(89)90508-4. [DOI] [PubMed] [Google Scholar]
  7. Chackalaparampil I., Shalloway D. Altered phosphorylation and activation of pp60c-src during fibroblast mitosis. Cell. 1988 Mar 25;52(6):801–810. doi: 10.1016/0092-8674(88)90422-9. [DOI] [PubMed] [Google Scholar]
  8. Chellappan S. P., Hiebert S., Mudryj M., Horowitz J. M., Nevins J. R. The E2F transcription factor is a cellular target for the RB protein. Cell. 1991 Jun 14;65(6):1053–1061. doi: 10.1016/0092-8674(91)90557-f. [DOI] [PubMed] [Google Scholar]
  9. Cleveland D. W. Autoregulated instability of tubulin mRNAs: a novel eukaryotic regulatory mechanism. Trends Biochem Sci. 1988 Sep;13(9):339–343. doi: 10.1016/0968-0004(88)90103-x. [DOI] [PubMed] [Google Scholar]
  10. Cleveland D. W., Yen T. J. Multiple determinants of eukaryotic mRNA stability. New Biol. 1989 Nov;1(2):121–126. [PubMed] [Google Scholar]
  11. Dawid I. B., Wellauer P. K. A reinvestigation of 5' leads to 3' polarity in 40S ribosomal RNA precursor of Xenopus laevis. Cell. 1976 Jul;8(3):443–448. doi: 10.1016/0092-8674(76)90157-4. [DOI] [PubMed] [Google Scholar]
  12. Elliott G. D., Meredith D. M. The herpes simplex virus type 1 tegument protein VP22 is encoded by gene UL49. J Gen Virol. 1992 Mar;73(Pt 3):723–726. doi: 10.1099/0022-1317-73-3-723. [DOI] [PubMed] [Google Scholar]
  13. Fenwick M. L., Clark J. Early and delayed shut-off of host protein synthesis in cells infected with herpes simplex virus. J Gen Virol. 1982 Jul;61(Pt 50):121–125. doi: 10.1099/0022-1317-61-1-121. [DOI] [PubMed] [Google Scholar]
  14. Fenwick M. L., Everett R. D. Transfer of UL41, the gene controlling virion-associated host cell shutoff, between different strains of herpes simplex virus. J Gen Virol. 1990 Feb;71(Pt 2):411–418. doi: 10.1099/0022-1317-71-2-411. [DOI] [PubMed] [Google Scholar]
  15. Fenwick M. L., McMenamin M. M. Early virion-associated suppression of cellular protein synthesis by herpes simplex virus is accompanied by inactivation of mRNA. J Gen Virol. 1984 Jul;65(Pt 7):1225–1228. doi: 10.1099/0022-1317-65-7-1225. [DOI] [PubMed] [Google Scholar]
  16. Fenwick M. L., Walker M. J. Suppression of the synthesis of cellular macromolecules by herpes simplex virus. J Gen Virol. 1978 Oct;41(1):37–51. doi: 10.1099/0022-1317-41-1-37. [DOI] [PubMed] [Google Scholar]
  17. Fenwick M., Morse L. S., Roizman B. Anatomy of herpes simplex virus DNA. XI. Apparent clustering of functions effecting rapid inhibition of host DNA and protein synthesis. J Virol. 1979 Feb;29(2):825–827. doi: 10.1128/jvi.29.2.825-827.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Frink R. J., Anderson K. P., Wagner E. K. Herpes simplex virus type 1 HindIII fragment L encodes spliced and complementary mRNA species. J Virol. 1981 Aug;39(2):559–572. doi: 10.1128/jvi.39.2.559-572.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Goldin A. L., Sandri-Goldin R. M., Levine M., Glorioso J. C. Cloning of herpes simplex virus type 1 sequences representing the whole genome. J Virol. 1981 Apr;38(1):50–58. doi: 10.1128/jvi.38.1.50-58.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Guo L. H., Stepień P. P., Tso J. Y., Brousseau R., Narang S., Thomas D. Y., Wu R. Synthesis of human insulin gene. VIII. Construction of expression vectors for fused proinsulin production in Escherichia coli. Gene. 1984 Jul-Aug;29(1-2):251–254. doi: 10.1016/0378-1119(84)90186-0. [DOI] [PubMed] [Google Scholar]
  21. Hentze M. W. Determinants and regulation of cytoplasmic mRNA stability in eukaryotic cells. Biochim Biophys Acta. 1991 Nov 11;1090(3):281–292. doi: 10.1016/0167-4781(91)90191-n. [DOI] [PubMed] [Google Scholar]
  22. Hill T. M., Sadler J. R., Betz J. L. Virion component of herpes simplex virus type 1 KOS interferes with early shutoff of host protein synthesis induced by herpes simplex virus type 2 186. J Virol. 1985 Oct;56(1):312–316. doi: 10.1128/jvi.56.1.312-316.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Johnson D. C., Feenstra V. Identification of a novel herpes simplex virus type 1-induced glycoprotein which complexes with gE and binds immunoglobulin. J Virol. 1987 Jul;61(7):2208–2216. doi: 10.1128/jvi.61.7.2208-2216.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Johnson D. C., Frame M. C., Ligas M. W., Cross A. M., Stow N. D. Herpes simplex virus immunoglobulin G Fc receptor activity depends on a complex of two viral glycoproteins, gE and gI. J Virol. 1988 Apr;62(4):1347–1354. doi: 10.1128/jvi.62.4.1347-1354.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kmiecik T. E., Shalloway D. Activation and suppression of pp60c-src transforming ability by mutation of its primary sites of tyrosine phosphorylation. Cell. 1987 Apr 10;49(1):65–73. doi: 10.1016/0092-8674(87)90756-2. [DOI] [PubMed] [Google Scholar]
  26. Kousoulas K. G., Bzik D. J., DeLuca N., Person S. The effect of ammonium chloride and tunicamycin on the glycoprotein content and infectivity of herpes simplex virus type 1. Virology. 1983 Mar;125(2):468–474. doi: 10.1016/0042-6822(83)90217-9. [DOI] [PubMed] [Google Scholar]
  27. Krikorian C. R., Read G. S. In vitro mRNA degradation system to study the virion host shutoff function of herpes simplex virus. J Virol. 1991 Jan;65(1):112–122. doi: 10.1128/jvi.65.1.112-122.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Krikorian C. R., Read G. S. Proteins associated with mRNA in cells infected with herpes simplex virus. Biochem Biophys Res Commun. 1989 Oct 16;164(1):355–361. doi: 10.1016/0006-291x(89)91726-9. [DOI] [PubMed] [Google Scholar]
  29. Kwong A. D., Frenkel N. Herpes simplex virus-infected cells contain a function(s) that destabilizes both host and viral mRNAs. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1926–1930. doi: 10.1073/pnas.84.7.1926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kwong A. D., Frenkel N. The herpes simplex virus virion host shutoff function. J Virol. 1989 Nov;63(11):4834–4839. doi: 10.1128/jvi.63.11.4834-4839.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kwong A. D., Kruper J. A., Frenkel N. Herpes simplex virus virion host shutoff function. J Virol. 1988 Mar;62(3):912–921. doi: 10.1128/jvi.62.3.912-921.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Liu F., Roizman B. Characterization of the protease and other products of amino-terminus-proximal cleavage of the herpes simplex virus 1 UL26 protein. J Virol. 1993 Mar;67(3):1300–1309. doi: 10.1128/jvi.67.3.1300-1309.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Marchetti M. E., Smith C. A., Schaffer P. A. A temperature-sensitive mutation in a herpes simplex virus type 1 gene required for viral DNA synthesis maps to coordinates 0.609 through 0.614 in UL. J Virol. 1988 Mar;62(3):715–721. doi: 10.1128/jvi.62.3.715-721.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. McGeoch D. J., Dalrymple M. A., Davison A. J., Dolan A., Frame M. C., McNab D., Perry L. J., Scott J. E., Taylor P. The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. J Gen Virol. 1988 Jul;69(Pt 7):1531–1574. doi: 10.1099/0022-1317-69-7-1531. [DOI] [PubMed] [Google Scholar]
  35. McLauchlan J., Clements J. B. Organization of the herpes simplex virus type 1 transcription unit encoding two early proteins with molecular weights of 140000 and 40000. J Gen Virol. 1983 May;64(Pt 5):997–1006. doi: 10.1099/0022-1317-64-5-997. [DOI] [PubMed] [Google Scholar]
  36. McLauchlan J., Phelan A., Loney C., Sandri-Goldin R. M., Clements J. B. Herpes simplex virus IE63 acts at the posttranscriptional level to stimulate viral mRNA 3' processing. J Virol. 1992 Dec;66(12):6939–6945. doi: 10.1128/jvi.66.12.6939-6945.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. O'Hare P., Hayward G. S. Expression of recombinant genes containing herpes simplex virus delayed-early and immediate-early regulatory regions and trans activation by herpesvirus infection. J Virol. 1984 Nov;52(2):522–531. doi: 10.1128/jvi.52.2.522-531.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Oroskar A. A., Read G. S. A mutant of herpes simplex virus type 1 exhibits increased stability of immediate-early (alpha) mRNAs. J Virol. 1987 Feb;61(2):604–606. doi: 10.1128/jvi.61.2.604-606.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Oroskar A. A., Read G. S. Control of mRNA stability by the virion host shutoff function of herpes simplex virus. J Virol. 1989 May;63(5):1897–1906. doi: 10.1128/jvi.63.5.1897-1906.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Peltz S. W., Brewer G., Bernstein P., Hart P. A., Ross J. Regulation of mRNA turnover in eukaryotic cells. Crit Rev Eukaryot Gene Expr. 1991;1(2):99–126. [PubMed] [Google Scholar]
  41. Read G. S., Frenkel N. Herpes simplex virus mutants defective in the virion-associated shutoff of host polypeptide synthesis and exhibiting abnormal synthesis of alpha (immediate early) viral polypeptides. J Virol. 1983 May;46(2):498–512. doi: 10.1128/jvi.46.2.498-512.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Read G. S., Sharp J. A., Summers W. C. In vitro and in vivo transcription initiation sites on the TK-encoding BamHI Q fragment of HSV-1 DNA. Virology. 1984 Oct 30;138(2):368–372. doi: 10.1016/0042-6822(84)90363-5. [DOI] [PubMed] [Google Scholar]
  43. Read G. S., Summers W. C. In vitro transcription of the thymidine kinase gene of herpes simplex virus. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5215–5219. doi: 10.1073/pnas.79.17.5215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Ross J. Messenger RNA turnover in eukaryotic cells. Mol Biol Med. 1988 Feb;5(1):1–14. [PubMed] [Google Scholar]
  45. Sandri-Goldin R. M., Levine M., Glorioso J. C. Method for induction of mutations in physically defined regions of the herpes simplex virus genome. J Virol. 1981 Apr;38(1):41–49. doi: 10.1128/jvi.38.1.41-49.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sandri-Goldin R. M., Mendoza G. E. A herpesvirus regulatory protein appears to act post-transcriptionally by affecting mRNA processing. Genes Dev. 1992 May;6(5):848–863. doi: 10.1101/gad.6.5.848. [DOI] [PubMed] [Google Scholar]
  47. Schek N., Bachenheimer S. L. Degradation of cellular mRNAs induced by a virion-associated factor during herpes simplex virus infection of Vero cells. J Virol. 1985 Sep;55(3):601–610. doi: 10.1128/jvi.55.3.601-610.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Smibert C. A., Johnson D. C., Smiley J. R. Identification and characterization of the virion-induced host shutoff product of herpes simplex virus gene UL41. J Gen Virol. 1992 Feb;73(Pt 2):467–470. doi: 10.1099/0022-1317-73-2-467. [DOI] [PubMed] [Google Scholar]
  49. Strom T., Frenkel N. Effects of herpes simplex virus on mRNA stability. J Virol. 1987 Jul;61(7):2198–2207. doi: 10.1128/jvi.61.7.2198-2207.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Tso J. Y., Sun X. H., Kao T. H., Reece K. S., Wu R. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucleic Acids Res. 1985 Apr 11;13(7):2485–2502. doi: 10.1093/nar/13.7.2485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Yaciuk P., Moran E. Analysis with specific polyclonal antiserum indicates that the E1A-associated 300-kDa product is a stable nuclear phosphoprotein that undergoes cell cycle phase-specific modification. Mol Cell Biol. 1991 Nov;11(11):5389–5397. doi: 10.1128/mcb.11.11.5389. [DOI] [PMC free article] [PubMed] [Google Scholar]

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