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. 1975 Jun;15(6):1348–1356. doi: 10.1128/jvi.15.6.1348-1356.1975

Both NS and L proteins are required for in vitro RNA synthesis by vesicular stomatitis virus.

S U Emerson, Y Yu
PMCID: PMC354602  PMID: 167189

Abstract

Vesicular stomatitis virions, Indiana serotype, were solubilized with high salt solubilizer and separated by ultracentrifugation into a supernatant fraction containing L, G, NS, and M proteins and pellet fraction containing the RNA complexed with N protein. NS protein was purified from the supernatnat fluid by sequential chromatography on phosphocellulose and diethylaminoethyl cellulose columns. The purified NS protein was assayed in a standard transcription system in combination with purified L protein and purified template (pellet fraction) prepared by renografin or CsCl banding. Results of the polymerase assays indicate that both L and NS proteins are required to reconstitute transcription activity with a highly purified template composed of only RNA and N protein. The NS protein polymerase activity is destroyed by trypsin but withstands 90 C temperatures for 10 min. Cytoplasmic NS protein can substitute for virion NS protein in the in vitro transcription assay.

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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. Banerjee A. K., Rhodes D. P. In vitro synthesis of RNA that contains polyadenylate by virion-associated RNA polymerase of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3566–3570. doi: 10.1073/pnas.70.12.3566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bishop D. H. Complete transcription by the transcriptase of vesicular stomatitis virus. J Virol. 1971 Apr;7(4):486–490. doi: 10.1128/jvi.7.4.486-490.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bishop D. H., Emerson S. U., Flamand A. Reconstitution of infectivity and transcriptase activity of homologous and heterologous viruses: vesicular stomatitis (Indiana serotype), Chandipura, vesicular stomatitis (New Jersey serotype), and Cocal viruses. J Virol. 1974 Jul;14(1):139–144. doi: 10.1128/jvi.14.1.139-144.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Bishop D. H., Roy P. Properties of the product synthesized by vesicular stomatitis virus particles. J Mol Biol. 1971 Jun 28;58(3):799–814. doi: 10.1016/0022-2836(71)90041-6. [DOI] [PubMed] [Google Scholar]
  7. Ehrenfeld E. Polyadenylation of vesicular stomatitis virus mRNA. J Virol. 1974 May;13(5):1055–1060. doi: 10.1128/jvi.13.5.1055-1060.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Galet H., Prevec L. Polyadenylate synthesis by extracts from L cells infected with vesicular stomatitis virus. Nat New Biol. 1973 Jun 13;243(128):200–203. doi: 10.1038/newbio243200a0. [DOI] [PubMed] [Google Scholar]
  11. Galet H., Shedlarski J. G., Jr, Prevec L. Ribonucleic acid polymerase induced in L-cells infected with vesicular stomatitis virus. Can J Biochem. 1973 May;51(5):721–729. doi: 10.1139/o73-089. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. 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]
  15. 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]
  16. Kang C. Y., Prevec L. Proteins of vesicular stomatitis virus. 3. Intracellular synthesis and extracellular appearance of virus-specific proteins. Virology. 1971 Dec;46(3):678–690. doi: 10.1016/0042-6822(71)90070-5. [DOI] [PubMed] [Google Scholar]
  17. Lafay F. Etude des mutants thermosensibles du Virus de la Stomatite Vésiculaire (VSV). Classification de quelques mutants d'après des critères de fonctionnement. C R Acad Sci Hebd Seances Acad Sci D. 1969 May 12;268(19):2385–2388. [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. Penman S., Scherrer K., Becker Y., Darnell J. E. POLYRIBOSOMES IN NORMAL AND POLIOVIRUS-INFECTED HELA CELLS AND THEIR RELATIONSHIP TO MESSENGER-RNA. Proc Natl Acad Sci U S A. 1963 May;49(5):654–662. doi: 10.1073/pnas.49.5.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Printz P., Wagner R. R. Temperature-sensitive mutants of vesicular stomatitis virus: synthesis of virus-specific proteins. J Virol. 1971 May;7(5):651–662. doi: 10.1128/jvi.7.5.651-662.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Villarreal L. P., Holland J. J. Synthesis of poly(A) in vitro by purified virions of vesicular stomatitis virus. Nat New Biol. 1973 Nov 7;246(149):17–19. doi: 10.1038/newbio246017a0. [DOI] [PubMed] [Google Scholar]
  25. Wagner R. R., Kiley M. P., Snyder R. M., Schnaitman C. A. Cytoplasmic compartmentalization of the protein and ribonucleic acid species of vesicular stomatitis virus. J Virol. 1972 Apr;9(4):672–683. doi: 10.1128/jvi.9.4.672-683.1972. [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., Snyder R. M., Yamazaki S. Proteins of vesicular stomatitis virus: kinetics and cellular sites of synthesis. J Virol. 1970 May;5(5):548–558. doi: 10.1128/jvi.5.5.548-558.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wong P. K., Holloway A. F., Cormack D. V. Characterization of three complementation groups of vesicular stomatitis virus. Virology. 1972 Dec;50(3):829–840. doi: 10.1016/0042-6822(72)90437-0. [DOI] [PubMed] [Google Scholar]

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