Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1979 Jul;31(1):8–15. doi: 10.1128/jvi.31.1.8-15.1979

Proposed replicative role of the NS polypeptide of vesicular stomatitis virus: structural analysis of an electrophoretic variant.

J A Lesnaw, L R Dickson, R H Curry
PMCID: PMC353416  PMID: 228061

Abstract

The structural lesion in the temperature-sensitive mutant E1 of the New Jersey serotype of vesicular stomatitis virus has been assigned to the NS protein. Although the packaged wild-type and mutant NS proteins were similarly phosphorylated, the mutant NS protein migrated faster than the wild-type NS protein in polyacrylamide slab gels electrophoresed in the presence of sodium dodecyl sulfate. The resolution appears to be the result of conformational rather than size differences since the two proteins comigrated in polyacrylamide gels which contained 4 M urea in addition to sodium dodecyl sulfate. Peptide maps, obtained by limited proteolysis of 32P-labeled wild-type and mutant NS proteins with Staphylococcus aureus V8 protease and papain, revealed striking differences which suggested that the mutant alteration could involve an aspartic or glutamic acid residue. Since NS proteins obtained from naturally occurring revertants of E1 were indistinguishable from the wild-type protein in all of these analyses, the structural alteration in the mutant NS protein correlates with the functional lesion. Because E1 is defective in the RNA replication pathway at the restrictive temperature, a replicative role is proposed for the NS protein.

Full text

PDF
8

Images in this article

9Image
on p.9
10Image
on p.10
11Image
on p.11
12Image
on p.12
13Image
on p.13
13Image
on p.13
14Image
on p.14

Selected References

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

  1. Baltimore D. Expression of animal virus genomes. Bacteriol Rev. 1971 Sep;35(3):235–241. doi: 10.1128/br.35.3.235-241.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  4. Clinton G. M., Burge B. W., Huang A. S. Effects of phosphorylation and pH on the association of NS protein with vesicular stomatitis virus cores. J Virol. 1978 Aug;27(2):340–346. doi: 10.1128/jvi.27.2.340-346.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Drapeau G. R., Boily Y., Houmard J. Purification and properties of an extracellular protease of Staphylococcus aureus. J Biol Chem. 1972 Oct 25;247(20):6720–6726. [PubMed] [Google Scholar]
  6. 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]
  7. Huang A. S., Manders E. K. Ribonucleic acid synthesis of vesicular stomatitis virus. IV. Transcription by standard virus in the presence of defective interfering particles. J Virol. 1972 Jun;9(6):909–916. doi: 10.1128/jvi.9.6.909-916.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Imblum R. L., Wagner R. R. Inhibition of viral transcriptase by immunoglobulin directed against the nucleocapsid NS protein of vesicular stomatitis virus. J Virol. 1975 Jun;15(6):1357–1366. doi: 10.1128/jvi.15.6.1357-1366.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Imblum R. L., Wagner R. R. Protein kinase and phosphoproteins of vesicular stomatitis virus. J Virol. 1974 Jan;13(1):113–124. doi: 10.1128/jvi.13.1.113-124.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. KIMMEL J. R., SMITH E. L. The properties of papain. Adv Enzymol Relat Subj Biochem. 1957;19:267–334. doi: 10.1002/9780470122648.ch4. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Lesnaw J. A., Dickson L. R. In vitro functional analysis of a temperature-sensitive mutant of vesicular stomatitis virus, New Jersey serotype, defective in transcription. Virology. 1978 Nov;91(1):51–59. doi: 10.1016/0042-6822(78)90354-9. [DOI] [PubMed] [Google Scholar]
  13. Lesnaw J. A., Reichmann M. E. RNA synthesis by temperature-sensitive mutants of vesicular stomatitis virus, New Jersey serotype. Virology. 1975 Feb;63(2):492–504. doi: 10.1016/0042-6822(75)90322-0. [DOI] [PubMed] [Google Scholar]
  14. MACPHERSON I., STOKER M. Polyoma transformation of hamster cell clones--an investigation of genetic factors affecting cell competence. Virology. 1962 Feb;16:147–151. doi: 10.1016/0042-6822(62)90290-8. [DOI] [PubMed] [Google Scholar]
  15. Marcus P. I., Engelhardt D. L., Hunt J. M., Sekellick M. J. Interferon action: inhibition of vesicular stomatitis virus RNA synthesis induced by virion-bound polymerase. Science. 1971 Nov 5;174(4009):593–598. doi: 10.1126/science.174.4009.593. [DOI] [PubMed] [Google Scholar]
  16. Moyer S. A., Summers D. F. Phosphorylation of vesicular stomatitis virus in vivo and in vitro. J Virol. 1974 Feb;13(2):455–465. doi: 10.1128/jvi.13.2.455-465.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Perlman S. M., Huang A. S. RNA synthesis of vesicular stomatitis virus. V. Interactions between transcription and replication. J Virol. 1973 Dec;12(6):1395–1400. doi: 10.1128/jvi.12.6.1395-1400.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Perrault J., Holland J. J. Absence of transcriptase activity or transcription-inhibiting ability in defective interfering particles of vesicular stomatitis virus. Virology. 1972 Oct;50(1):150–170. [PubMed] [Google Scholar]
  19. Pringle C. R., Duncan I. B., Stevenson M. Isolation and characterization of temperature-sensitive mutants of vesicular stomatitis virus, New Jersey serotype. J Virol. 1971 Dec;8(6):836–841. doi: 10.1128/jvi.8.6.836-841.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sokol F., Clark H. F. Phosphoproteins, structural components of rhabdoviruses. Virology. 1973 Mar;52(1):246–263. doi: 10.1016/0042-6822(73)90413-3. [DOI] [PubMed] [Google Scholar]
  21. Sokol F., Tan K. B., McFalls M. L., Madore P. Phosphate acceptor amino acid residues in structural proteins of rhabdoviruses. J Virol. 1974 Jul;14(1):145–151. doi: 10.1128/jvi.14.1.145-151.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  23. Wertz G. W., Levine M. RNA synthesis by vesicular stomatitis virus and a small plaque mutant: effects of cycloheximide. J Virol. 1973 Aug;12(2):253–264. doi: 10.1128/jvi.12.2.253-264.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES