Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1982 Oct;44(1):189–198. doi: 10.1128/jvi.44.1.189-198.1982

Evidence that vesicular stomatitis virus produces double-stranded RNA that inhibits protein synthesis in a reticulocyte lysate.

J R Thomas, R R Wagner
PMCID: PMC256252  PMID: 6183445

Abstract

Cell-free protein synthesis by reticulocyte lysates was inhibited by a polyadenylated RNA fraction extracted from HeLa cells infected with vesicular stomatitis virus (VSV) but not by polyadenylated RNA from mock-infected HeLa cells. A similar inhibitor of cell-free protein synthesis was found in a polyadenylated fraction of RNA transcribed in vitro by VSV but not in untranscribed nucleocapsids. Fractionation of the VSV transcription product showed that the translation inhibitor segregated with nucleocapsids containing newly transcribed polyadenylated or non-polyadenylated RNA, as determined by oligodeoxythymidylate-cellulose chromatography. The inhibitors present in VSV-infected HeLa cells and in VSV in vitro transcripts both appeared to be double-stranded RNA, as judged by the characteristics for inhibition of reticulocyte cell-free protein synthesis described by Hunter et al. (J. Biol. Chem. 250:409-417, 1975). The double-stranded nature of the VSV RNA inhibitor was supported by the finding that the translational inhibitory effect was inactivated by melting the inhibitor in the absence of salt and by micrococcal nuclease.

Full text

PDF
189

Images in this article

191Image
on p.191
194Image
on p.194

Selected References

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

  1. Baglioni C., Lenz J. R., Maroney P. A., Weber L. A. Effect of double-stranded RNA associated with viral messenger RNA on in vitro protein synthesis. Biochemistry. 1978 Aug 8;17(16):3257–3262. doi: 10.1021/bi00609a013. [DOI] [PubMed] [Google Scholar]
  2. Barenholz Y., Moore N. F., Wagner R. R. Enveloped viruses as model membrane systems: microviscosity of vesicular stomatitis virus and host cell membranes. Biochemistry. 1976 Aug 10;15(16):3563–3570. doi: 10.1021/bi00661a026. [DOI] [PubMed] [Google Scholar]
  3. Brown G. E., Lebleu B., Kawakita M., Shaila S., Sen G. C., Lengyel P. Increased endonuclease activity in an extract from mouse Ehrlich ascites tumor cells which had been treated with a partially purified interferon preparation: dependence of double-stranded RNA;. Biochem Biophys Res Commun. 1976 Mar 8;69(1):114–122. doi: 10.1016/s0006-291x(76)80280-x. [DOI] [PubMed] [Google Scholar]
  4. Carroll A. R., Wagner R. R. Inhibition of transcription by immunoglobulins directed against the ribonucleoprotein of homotypic and heterotypic vesicular stomatitis viruses. J Virol. 1978 Feb;25(2):675–684. doi: 10.1128/jvi.25.2.675-684.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Celma M. L., Ehrenfeld E. Effect of poliovirus double-stranded RNA on viral and host-cell protein synthesis. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2440–2444. doi: 10.1073/pnas.71.6.2440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clemens M. J., Safer B., Merrick W. C., Anderson W. F., London I. M. Inhibition of protein synthesis in rabbit reticulocyte lysates by double-stranded RNA and oxidized glutathione: indirect mode of action on polypeptide chain initiation. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1286–1290. doi: 10.1073/pnas.72.4.1286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Colby C., Morgan M. J. Interferon induction and action. Annu Rev Microbiol. 1971;25:333–360. doi: 10.1146/annurev.mi.25.100171.002001. [DOI] [PubMed] [Google Scholar]
  8. Darnbrough C., Hunt T., Jackson R. J. A complex between met-tRNA F and native 40S subunits in reticulocyte lysates and its disappearance during incubation with double-stranded RNA. Biochem Biophys Res Commun. 1972 Sep 26;48(6):1556–1564. doi: 10.1016/0006-291x(72)90891-1. [DOI] [PubMed] [Google Scholar]
  9. Dubovi E. J., Wagner R. R. Spatial relationships of the proteins of vesicular stomatitis virus: induction of reversible oligomers by cleavable protein cross-linkers and oxidation. J Virol. 1977 May;22(2):500–509. doi: 10.1128/jvi.22.2.500-509.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ehrenfeld E., Hunt T. Double-stranded poliovirus RNA inhibits initiation of protein synthesis by reticulocyte lysates. Proc Natl Acad Sci U S A. 1971 May;68(5):1075–1078. doi: 10.1073/pnas.68.5.1075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Farrell P. J., Balkow K., Hunt T., Jackson R. J., Trachsel H. Phosphorylation of initiation factor elF-2 and the control of reticulocyte protein synthesis. Cell. 1977 May;11(1):187–200. doi: 10.1016/0092-8674(77)90330-0. [DOI] [PubMed] [Google Scholar]
  12. Graziadei W. D., 3rd, Lengyel P. Translation of in vitro synthesized reovirus messenger RNAs into proteins of the size of reovirus capsid proteins in a mouse L cell extract. Biochem Biophys Res Commun. 1972 Mar 10;46(5):1816–1823. doi: 10.1016/0006-291x(72)90056-3. [DOI] [PubMed] [Google Scholar]
  13. Hunter T., Hunt T., Jackson R. J., Robertson H. D. The characteristics of inhibition of protein synthesis by double-stranded ribonucleic acid in reticulocyte lysates. J Biol Chem. 1975 Jan 25;250(2):409–417. [PubMed] [Google Scholar]
  14. Kaempfer R., Kaufman J. Inhibition of cellular protein synthesis by double-stranded RNA: inactivation of an initiation factor. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1222–1226. doi: 10.1073/pnas.70.4.1222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. McAllister P. E., Wagner R. R. Differential inhibition of host protein synthesis in L cells infected with RNA - temperature-sensitive mutants of vesicular stomatitis virus. J Virol. 1976 May;18(2):550–558. doi: 10.1128/jvi.18.2.550-558.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McDowell M. J., Joklik W. K., Villa-Komaroff L., Lodish H. F. Translation of reovirus messenger RNAs synthetesized in vitro into reovirus polypeptides by several mammalian cell-free extracts. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2649–2653. doi: 10.1073/pnas.69.9.2649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Moyer S. A., Banerjee A. K. Messenger RNA species synthesized in vitro by the virion-associated RNA polymerase of vesicular stomatitis virus. Cell. 1975 Jan;4(1):37–43. doi: 10.1016/0092-8674(75)90131-2. [DOI] [PubMed] [Google Scholar]
  20. Nilsen T. W., Maroney P. A., Baglioni C. Double-stranded RNA causes synthesis of 2',5'-oligo(A) and degradation of messenger RNA in interferon-treated cells. J Biol Chem. 1981 Aug 10;256(15):7806–7811. [PubMed] [Google Scholar]
  21. Nilsen T. W., Wood D. L., Baglioni C. Cross-linking of viral RNA by 4'-aminomethyl-4,5',8-trimethylpsoralen in HeLa cells infected with encephalomyocarditis virus and the tsG114 mutant of vesicular stomatitis virus. Virology. 1981 Feb;109(1):82–93. doi: 10.1016/0042-6822(81)90473-6. [DOI] [PubMed] [Google Scholar]
  22. Nuss D. L., Koch G. Translation of individual host mRNA's in MPC-11 cells is differentially suppressed after infection by vesicular stomatitis virus. J Virol. 1976 Aug;19(2):572–578. doi: 10.1128/jvi.19.2.572-578.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Palmiter R. D. Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid. Biochemistry. 1974 Aug 13;13(17):3606–3615. doi: 10.1021/bi00714a032. [DOI] [PubMed] [Google Scholar]
  24. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  25. Roberts W. K., Clemens M. J., Kerr I. M. Interferon-induced inhibition of protein synthesis in L-cell extracts: an ATP-dependent step in the activation of an inhibitor by double-stranded RNA. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3136–3140. doi: 10.1073/pnas.73.9.3136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Robertson H. D., Mathews M. B. Double-stranded RNA as an inhibitor of protein synthesis and as a substrate for a nuclease in extracts of Krebs II ascites cells. Proc Natl Acad Sci U S A. 1973 Jan;70(1):225–229. doi: 10.1073/pnas.70.1.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. WAGNER R. R., LEVEY A. H., SNYDER R. M., RATCLIFF G. A., Jr, HYATT D. F. BIOLOGIC PROPERTIES OF TWO PLAQUE VARIANTS OF VESICULAR STOMATITIS VIRUS (INDIANA SEROTYPE). J Immunol. 1963 Jul;91:112–122. [PubMed] [Google Scholar]

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

RESOURCES