Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1996 Apr;70(4):2411–2419. doi: 10.1128/jvi.70.4.2411-2419.1996

The virion host shutoff protein of herpes simplex virus type 1: messenger ribonucleolytic activity in vitro.

B D Zelus 1, R S Stewart 1, J Ross 1
PMCID: PMC190084  PMID: 8642669

Abstract

Shortly after tissue culture cells are infected with herpes simplex virus (HSV) type 1 or 2, the rate of host protein synthesis decreases 5- to 10-fold and most host mRNAs are degraded. mRNA destabilization is triggered by the virion host shutoff (vhs) protein, a virus encoded, 58-kDa protein located in the virion tegument. To determine whether it can function as a messenger RNase (mRNase), the capacity of vhs protein to degrade RNA in vitro in absence of host cell components was assessed. Two sources of vhs protein were used in these assays: crude extract from virions or protein translated in a reticulocyte-free system. In each case, wild-type but not mutant vhs protein degraded various RNA substrates. Preincubation with anti-vhs antibody blocked RNase activity. These studies do not prove that vhs protein on its own is an mRNase but do demonstrate that the protein, either on its own or in conjunction with another factor(s), has the biochemical property of an mRNase, consistent with its role in infected cells.

Full Text

The Full Text of this article is available as a PDF (462.4 KB).

Selected References

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

  1. Bandyopadhyay R., Coutts M., Krowczynska A., Brawerman G. Nuclease activity associated with mammalian mRNA in its native state: possible basis for selectivity in mRNA decay. Mol Cell Biol. 1990 May;10(5):2060–2069. doi: 10.1128/mcb.10.5.2060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Becker Y., Tavor E., Asher Y., Berkowitz C., Moyal M. Effect of herpes simplex virus type-1 UL41 gene on the stability of mRNA from the cellular genes: beta-actin, fibronectin, glucose transporter-1, and docking protein, and on virus intraperitoneal pathogenicity to newborn mice. Virus Genes. 1993 Jun;7(2):133–143. doi: 10.1007/BF01702393. [DOI] [PubMed] [Google Scholar]
  3. Bernstein P. L., Herrick D. J., Prokipcak R. D., Ross J. Control of c-myc mRNA half-life in vitro by a protein capable of binding to a coding region stability determinant. Genes Dev. 1992 Apr;6(4):642–654. doi: 10.1101/gad.6.4.642. [DOI] [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. Caruccio N., Ross J. Purification of a human polyribosome-associated 3' to 5' exoribonuclease. J Biol Chem. 1994 Dec 16;269(50):31814–31821. [PubMed] [Google Scholar]
  6. Dompenciel R. E., Garnepudi V. R., Schoenberg D. R. Purification and characterization of an estrogen-regulated Xenopus liver polysomal nuclease involved in the selective destabilization of albumin mRNA. J Biol Chem. 1995 Mar 17;270(11):6108–6118. doi: 10.1074/jbc.270.11.6108. [DOI] [PubMed] [Google Scholar]
  7. Eichler D. C., Craig N. Processing of eukaryotic ribosomal RNA. Prog Nucleic Acid Res Mol Biol. 1994;49:197–239. doi: 10.1016/s0079-6603(08)60051-3. [DOI] [PubMed] [Google Scholar]
  8. Eichler D. C., Eales S. J. Isolation and characterization of a single-stranded specific endoribonuclease from Ehrlich cell nucleoli. J Biol Chem. 1982 Dec 10;257(23):14384–14389. [PubMed] [Google Scholar]
  9. Eichler D. C., Eales S. J. Purification and properties of a novel nucleolar exoribonuclease capable of degrading both single-stranded and double-stranded RNA. Biochemistry. 1985 Jan 29;24(3):686–691. doi: 10.1021/bi00324a022. [DOI] [PubMed] [Google Scholar]
  10. Eichler D. C., Liberatore J. A., Shumard C. M. Selection of a preribosomal RNA processing site by a nucleolar endoribonuclease involves formation of a stable complex. Nucleic Acids Res. 1993 Dec 11;21(24):5775–5781. doi: 10.1093/nar/21.24.5775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Elroy-Stein O., Fuerst T. R., Moss B. Cap-independent translation of mRNA conferred by encephalomyocarditis virus 5' sequence improves the performance of the vaccinia virus/bacteriophage T7 hybrid expression system. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6126–6130. doi: 10.1073/pnas.86.16.6126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  13. Fenwick M. L., Clark J. The effect of cycloheximide on the accumulation and stability of functional alpha-mRNA in cells infected with herpes simplex virus. J Gen Virol. 1983 Sep;64(Pt 9):1955–1963. doi: 10.1099/0022-1317-64-9-1955. [DOI] [PubMed] [Google Scholar]
  14. Fenwick M. L., Everett R. D. Inactivation of the shutoff gene (UL41) of herpes simplex virus types 1 and 2. J Gen Virol. 1990 Dec;71(Pt 12):2961–2967. doi: 10.1099/0022-1317-71-12-2961. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Gorman C., Padmanabhan R., Howard B. H. High efficiency DNA-mediated transformation of primate cells. Science. 1983 Aug 5;221(4610):551–553. doi: 10.1126/science.6306768. [DOI] [PubMed] [Google Scholar]
  18. Herrick D. J., Ross J. The half-life of c-myc mRNA in growing and serum-stimulated cells: influence of the coding and 3' untranslated regions and role of ribosome translocation. Mol Cell Biol. 1994 Mar;14(3):2119–2128. doi: 10.1128/mcb.14.3.2119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Jones F. E., Smibert C. A., Smiley J. R. Mutational analysis of the herpes simplex virus virion host shutoff protein: evidence that vhs functions in the absence of other viral proteins. J Virol. 1995 Aug;69(8):4863–4871. doi: 10.1128/jvi.69.8.4863-4871.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Krieg P. A., Melton D. A. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Res. 1984 Sep 25;12(18):7057–7070. doi: 10.1093/nar/12.18.7057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Lang K. M., Spritz R. A. In vitro splicing pathways of pre-mRNAs containing multiple intervening sequences? Mol Cell Biol. 1987 Oct;7(10):3428–3437. doi: 10.1128/mcb.7.10.3428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Masters D. B., Griggs C. T., Berde C. B. High sensitivity quantification of RNA from gels and autoradiograms with affordable optical scanning. Biotechniques. 1992 Jun;12(6):902-6, 908-11. [PubMed] [Google Scholar]
  26. McLauchlan J., Rixon F. J. Characterization of enveloped tegument structures (L particles) produced by alphaherpesviruses: integrity of the tegument does not depend on the presence of capsid or envelope. J Gen Virol. 1992 Feb;73(Pt 2):269–276. doi: 10.1099/0022-1317-73-2-269. [DOI] [PubMed] [Google Scholar]
  27. Muhlrad D., Decker C. J., Parker R. Deadenylation of the unstable mRNA encoded by the yeast MFA2 gene leads to decapping followed by 5'-->3' digestion of the transcript. Genes Dev. 1994 Apr 1;8(7):855–866. doi: 10.1101/gad.8.7.855. [DOI] [PubMed] [Google Scholar]
  28. Nakai H., Maxwell I. H., Pizer L. I. Herpesvirus infection alters the steady-state levels of cellular polyadenylated RNA in polyoma virus-transformed BHK cells. J Virol. 1982 Jun;42(3):1131–1134. doi: 10.1128/jvi.42.3.1131-1134.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Pak A. S., Everly D. N., Knight K., Read G. S. The virion host shutoff protein of herpes simplex virus inhibits reporter gene expression in the absence of other viral gene products. Virology. 1995 Aug 20;211(2):491–506. doi: 10.1006/viro.1995.1431. [DOI] [PubMed] [Google Scholar]
  31. Preston C. M., Frame M. C., Campbell M. E. A complex formed between cell components and an HSV structural polypeptide binds to a viral immediate early gene regulatory DNA sequence. Cell. 1988 Feb 12;52(3):425–434. doi: 10.1016/s0092-8674(88)80035-7. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Read G. S., Karr B. M., Knight K. 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. J Virol. 1993 Dec;67(12):7149–7160. doi: 10.1128/jvi.67.12.7149-7160.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ross J., Kobs G., Brewer G., Peltz S. W. Properties of the exonuclease activity that degrades H4 histone mRNA. J Biol Chem. 1987 Jul 5;262(19):9374–9381. [PubMed] [Google Scholar]
  35. Ross J., Kobs G. H4 histone messenger RNA decay in cell-free extracts initiates at or near the 3' terminus and proceeds 3' to 5'. J Mol Biol. 1986 Apr 20;188(4):579–593. doi: 10.1016/s0022-2836(86)80008-0. [DOI] [PubMed] [Google Scholar]
  36. Ross J. mRNA stability in mammalian cells. Microbiol Rev. 1995 Sep;59(3):423–450. doi: 10.1128/mr.59.3.423-450.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. 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]
  38. 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]
  39. Shumard C. M., Eichler D. C. Ribosomal RNA processing. Limited cleavages of mouse preribosomal RNA by a nucleolar endoribonuclease include the early +650 processing site. J Biol Chem. 1988 Dec 25;263(36):19346–19352. [PubMed] [Google Scholar]
  40. Shumard C. M., Torres C., Eichler D. C. In vitro processing at the 3'-terminal region of pre-18S rRNA by a nucleolar endoribonuclease. Mol Cell Biol. 1990 Aug;10(8):3868–3872. doi: 10.1128/mcb.10.8.3868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Silverstein S., Engelhardt D. L. Alterations in the protein synthetic apparatus of cells infected with herpes simplex virus. Virology. 1979 Jun;95(2):334–342. doi: 10.1016/0042-6822(79)90488-4. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. Smibert C. A., Popova B., Xiao P., Capone J. P., Smiley J. R. Herpes simplex virus VP16 forms a complex with the virion host shutoff protein vhs. J Virol. 1994 Apr;68(4):2339–2346. doi: 10.1128/jvi.68.4.2339-2346.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Smith I. L., Hardwicke M. A., Sandri-Goldin R. M. Evidence that the herpes simplex virus immediate early protein ICP27 acts post-transcriptionally during infection to regulate gene expression. Virology. 1992 Jan;186(1):74–86. doi: 10.1016/0042-6822(92)90062-t. [DOI] [PubMed] [Google Scholar]
  45. Sorenson C. M., Hart P. A., Ross J. Analysis of herpes simplex virus-induced mRNA destabilizing activity using an in vitro mRNA decay system. Nucleic Acids Res. 1991 Aug 25;19(16):4459–4465. doi: 10.1093/nar/19.16.4459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Stevens A., Maupin M. K. A 5'----3' exoribonuclease of Saccharomyces cerevisiae: size and novel substrate specificity. Arch Biochem Biophys. 1987 Feb 1;252(2):339–347. doi: 10.1016/0003-9861(87)90040-3. [DOI] [PubMed] [Google Scholar]
  47. 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]
  48. Visalli R. J., Brandt C. R. The HSV-1 UL45 18 kDa gene product is a true late protein and a component of the virion. Virus Res. 1993 Aug;29(2):167–178. doi: 10.1016/0168-1702(93)90057-t. [DOI] [PubMed] [Google Scholar]
  49. Zhou A., Hassel B. A., Silverman R. H. Expression cloning of 2-5A-dependent RNAase: a uniquely regulated mediator of interferon action. Cell. 1993 Mar 12;72(5):753–765. doi: 10.1016/0092-8674(93)90403-d. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES