Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1977 May;22(2):381–388. doi: 10.1128/jvi.22.2.381-388.1977

Temperature-dependent host range mutation in vesicular stomatitis virus affecting polypeptide L.

J F Szilágyi, C R Pringle, T M Macpherson
PMCID: PMC515729  PMID: 194060

Abstract

We established previously that the temperature-dependent host range mutant, td CE 3, of vesicular stomatitis virus (VSV) New Jersey possesses temperature-sensitive RNA transcriptase activity. In this paper, we describe dissociation and reconstitution experiments designed to determine which VSV polypeptide is affected by the td CE 3 mutation. Wild-type VSV New Jersey (ts+), the temperature-dependent host range mutant (td CE 3), and the revertant of this mutant (td CE/R1) were used. Transcribing nucleoprotein preparations, isolated from purified virus particles, were treated in the presence of digitonin with either 0.9 M LiCl to produce supernatants containing virtually only the L polypeptide or 2.0 M LiCl to produce ribonucleoprotein pellets containing only the polypeptides N and NS. Supernatant and pellet fractions synthesized either no or only trace amounts of RNA in vitro. Reconstitution of the supernatants with the pellets in all combinations at 31 degrees C restored much of the transcriptase activity of the transcribing nucleoprotein preparations. RNA synthesis occurred at 39 degrees C when the three pellets were reconstituted with wild-type and revertant supernatants. However, supernatant of the mutant td CE 3 reconstituted with any of the three pellets resulted in little or no detectable transcriptase activity at 39 degrees C. This implies that the polypeptide affected by the td CE 3 mutation is the L polypeptide.

Full text

PDF
381

Images in this article

384Image
on p.384

Selected References

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

  1. Baltimore D., Huang A. S., Stampfer M. Ribonucleic acid synthesis of vesicular stomatitis virus, II. An RNA polymerase in the virion. Proc Natl Acad Sci U S A. 1970 Jun;66(2):572–576. doi: 10.1073/pnas.66.2.572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bishop D. H., Roy P. Dissociation of vesicular stomatitis virus and relation of the virion proteins to the viral transcriptase. J Virol. 1972 Aug;10(2):234–243. doi: 10.1128/jvi.10.2.234-243.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bishop D. H., Roy P. Kinetics of RNA synthesis by vesicular stomatitis virus particles. J Mol Biol. 1971 May 14;57(3):513–527. doi: 10.1016/0022-2836(71)90106-9. [DOI] [PubMed] [Google Scholar]
  4. Brown F., Cartwright B., Crick J., Smale C. J. Infective virus substructure from vesicular stomatitis virus. J Virol. 1967 Apr;1(2):368–373. doi: 10.1128/jvi.1.2.368-373.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Emerson S. U., Wagner R. R. Dissociation and reconstitution of the transcriptase and template activities of vesicular stomatitis B and T virions. J Virol. 1972 Aug;10(2):297–309. doi: 10.1128/jvi.10.2.297-309.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Emerson S. U., Wagner R. R. L protein requirement for in vitro RNA synthesis by vesicular stomatitis virus. J Virol. 1973 Dec;12(6):1325–1335. doi: 10.1128/jvi.12.6.1325-1335.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Emerson S. U., Yu Y. Both NS and L proteins are required for in vitro RNA synthesis by vesicular stomatitis virus. J Virol. 1975 Jun;15(6):1348–1356. doi: 10.1128/jvi.15.6.1348-1356.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Huang A. S., Baltimore D., Stampfer M. Ribonucleic acid synthesis of vesicular stomatitis virus. 3. Multiple complementary messenger RNA molecules. Virology. 1970 Dec;42(4):946–957. doi: 10.1016/0042-6822(70)90343-0. [DOI] [PubMed] [Google Scholar]
  9. Huang A. S., Wagner R. R. Comparative sedimentation coefficients of RNA extracted from plaque-forming and defective particles of vesicular stomatitis virus. J Mol Biol. 1966 Dec 28;22(2):381–384. doi: 10.1016/0022-2836(66)90143-4. [DOI] [PubMed] [Google Scholar]
  10. Hunt D. M., Wagner R. R. Location of the transcription defect in group I temperature-sensitive mutants of vesicular stomatitis virus. J Virol. 1974 Jan;13(1):28–35. doi: 10.1128/jvi.13.1.28-35.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kang C. Y., Prevec L. Proteins of vesicular stomatitis virus. I. Polyacrylamide gel analysis of viral antigens. J Virol. 1969 Apr;3(4):404–413. doi: 10.1128/jvi.3.4.404-413.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Klenk H. D., Choppin P. W. Glycolipid content of vesicular stomatitis virus grown in baby hamster kidney cells. J Virol. 1971 Mar;7(3):416–417. doi: 10.1128/jvi.7.3.416-417.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Marsden H. S., Crombie I. K., Subak-Sharpe J. H. Control of protein synthesis in herpesvirus-infected cells: analysis of the polypeptides induced by wild type and sixteen temperature-sensitive mutants of HSV strain 17. J Gen Virol. 1976 Jun;31(3):347–372. doi: 10.1099/0022-1317-31-3-347. [DOI] [PubMed] [Google Scholar]
  15. McCrae M. A., Szilágyi J. F. Preparation and characterisation of a subviral particle of vaccinia virus containing the DNA-dependent RNA polymerase activity. Virology. 1975 Nov;68(1):234–244. doi: 10.1016/0042-6822(75)90164-6. [DOI] [PubMed] [Google Scholar]
  16. McSharry J. J., Wagner R. R. Lipid composition of purified vesicular stomatitis viruses. J Virol. 1971 Jan;7(1):59–70. doi: 10.1128/jvi.7.1.59-70.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mudd J. A., Summers D. F. Polysomal ribonucleic acid of vesicular stomatitis virus-infected HeLa cells. Virology. 1970 Dec;42(4):958–968. doi: 10.1016/0042-6822(70)90344-2. [DOI] [PubMed] [Google Scholar]
  18. Mudd J. A., Summers D. F. Protein synthesis in vesicular stomatitis virus-infected HeLa cells. Virology. 1970 Oct;42(2):328–340. doi: 10.1016/0042-6822(70)90277-1. [DOI] [PubMed] [Google Scholar]
  19. Naito S., Ishihama A. Function and structure of RNA polymerase from vesicular stomatitis virus. J Biol Chem. 1976 Jul 25;251(14):4307–4314. [PubMed] [Google Scholar]
  20. Obijeski J. F., Simpson R. W. Conditional lethal mutants of vesicular stomatitis virus. II. Synthesis of virus-specific polypeptides in nonpermissive cells infected with "RNA-" host-restricted mutants. Virology. 1974 Feb;57(2):369–377. doi: 10.1016/0042-6822(74)90176-7. [DOI] [PubMed] [Google Scholar]
  21. Pringle C. R. Conditional lethal mutants of vesicular stomatitis virus. Curr Top Microbiol Immunol. 1975;69:85–116. doi: 10.1007/978-3-642-50112-8_2. [DOI] [PubMed] [Google Scholar]
  22. Repik P., Flamand A., Bishop D. H. Synthesis of RNA by mutants of vesicular stomatitis virus (Indiana serotype) and the ability of wild-type VSV New Jersey to complement the VSV Indiana ts G I-114 transcription defect. J Virol. 1976 Oct;20(1):157–169. doi: 10.1128/jvi.20.1.157-169.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Szilágyi J. F., Pringle C. R. Effect of temperature-sensitive mutations on the virion-associated RNA transcriptase of vesicular stomatitis virus. J Mol Biol. 1972 Nov 14;71(2):281–291. doi: 10.1016/0022-2836(72)90351-8. [DOI] [PubMed] [Google Scholar]
  24. Szilágyi J. F., Pringle C. R. Virion trascriptase activity differences in host range mutants of vesicular stomatitis virus. J Virol. 1975 Oct;16(4):927–936. doi: 10.1128/jvi.16.4.927-936.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Szilágyi J. F., Uryvayev L. Isolation of an infectious ribonucleoprotein from vesicular stomatitis virus containing an active RNA transcriptase. J Virol. 1973 Feb;11(2):279–286. doi: 10.1128/jvi.11.2.279-286.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wagner R. R., Prevec L., Brown F., Summers D. F., Sokol F., MacLeod R. Classification of rhabdovirus proteins: a proposal. J Virol. 1972 Dec;10(6):1228–1230. doi: 10.1128/jvi.10.6.1228-1230.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wagner R. R., Schnaitman T. A., Snyder R. M. Structural proteins of vesicular stomatitis viruses. J Virol. 1969 Apr;3(4):395–403. doi: 10.1128/jvi.3.4.395-403.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wunner W. H., Pringle C. R. Protein synthesis in BHK21 cells infected with vesicular stomatitis virus. I. ts Mutants of the Indiana serotype. Virology. 1972 Apr;48(1):104–111. doi: 10.1016/0042-6822(72)90118-3. [DOI] [PubMed] [Google Scholar]

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

RESOURCES