Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1985 Jan;53(1):44–51. doi: 10.1128/jvi.53.1.44-51.1985

Assignment of the temperature-sensitive lesion in the replication mutant A1 of vesicular stomatitis virus to the N gene.

M D Marks, J Kennedy-Morrow, J A Lesnaw
PMCID: PMC254976  PMID: 2981361

Abstract

The replication defect in the temperature-sensitive mutant A1 of the New Jersey serotype (Hazelhurst subtype) of vesicular stomatitis virus was confirmed by the absence of intracellular nucleocapsids in infected cells incubated at the restrictive temperature. After preamplification, the relative yield of the A1 N protein accumulated intracellularly after 1 h of incubation at the restrictive temperature was decreased by 50% that of the wild-type or revertant A1 N protein. This difference was not as apparent in pulse-chase experiments. The functional lesion in A1 was correlated with a structural alteration in the N protein on the basis of the thermolability of the template activity of the A1 N protein-RNA complex in in vitro transcription reactions and the covariance of this phenotype with the temperature-sensitive phenotype in a spontaneous A1 revertant. This correlation was consistent with a direct role of the N protein in replication and allowed the assignment of the N gene to complementation group A.

Full text

PDF
44

Images in this article

47Image
on p.47
48Image
on p.48

Selected References

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

  1. Ball L. A., Wertz G. W. VSV RNA synthesis: how can you be positive? Cell. 1981 Oct;26(2 Pt 2):143–144. doi: 10.1016/0092-8674(81)90297-x. [DOI] [PubMed] [Google Scholar]
  2. Blumberg B. M., Leppert M., Kolakofsky D. Interaction of VSV leader RNA and nucleocapsid protein may control VSV genome replication. Cell. 1981 Mar;23(3):837–845. doi: 10.1016/0092-8674(81)90448-7. [DOI] [PubMed] [Google Scholar]
  3. Evans D., Pringle C. R., Szilágyi J. F. Temperature-sensitive mutants of complementation group E of vesicular stomatitis virus New Jersey serotype possess altered NS polypeptides. J Virol. 1979 Aug;31(2):325–333. doi: 10.1128/jvi.31.2.325-333.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Isle H. D., Emerson S. U. Use of a hybrid infectivity assay to analyze primary transcription of temperature-sensitive mutants of the New Jersey serotype of vesicular stomatitis virus. J Virol. 1982 Jul;43(1):37–40. doi: 10.1128/jvi.43.1.37-40.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kennedy-Morrow J., Lesnaw J. A. Structural and functional characterization of the RNA-positive complementation groups, C and D, of the New Jersey serotype of vesicular stomatitis virus: assignment of the M gene to the C complementation group. Virology. 1984 Jan 15;132(1):38–52. doi: 10.1016/0042-6822(84)90089-8. [DOI] [PubMed] [Google Scholar]
  6. Kingsbury D. W. The molecular biology of paramyxoviruses. Med Microbiol Immunol. 1974;160(2-3):73–83. doi: 10.1007/BF02121714. [DOI] [PubMed] [Google Scholar]
  7. Knipe D., Lodish H. F., Baltimore D. Analysis of the defects of temperature-sensitive mutants of vesicular stomatitis virus: intracellular degradation of specific viral proteins. J Virol. 1977 Mar;21(3):1140–1148. doi: 10.1128/jvi.21.3.1140-1148.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Lesnaw J. A., Dickson L. R., Curry R. H. Proposed replicative role of the NS polypeptide of vesicular stomatitis virus: structural analysis of an electrophoretic variant. J Virol. 1979 Jul;31(1):8–15. doi: 10.1128/jvi.31.1.8-15.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Little S. P., Huang A. S. Shedding of the glycoprotein from vesicular stomatitis virus-infected cells. J Virol. 1978 Aug;27(2):330–339. doi: 10.1128/jvi.27.2.330-339.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Patton J. T., Davis N. L., Wertz G. W. N protein alone satisfies the requirement for protein synthesis during RNA replication of vesicular stomatitis virus. J Virol. 1984 Feb;49(2):303–309. doi: 10.1128/jvi.49.2.303-309.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Pringle C. R., Devine V., Wilkie M., Preston C. M., Dolan A., McGeoch D. J. Enhanced mutability associated with a temperature-sensitive mutant of vesicular stomatitis virus. J Virol. 1981 Aug;39(2):377–389. doi: 10.1128/jvi.39.2.377-389.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Pringle C. R. The genetics of vesiculoviruses. Arch Virol. 1982;72(1-2):1–34. doi: 10.1007/BF01314447. [DOI] [PubMed] [Google Scholar]
  19. Szilágyi J. F., Pringle C. R. Effect of temperature-sensitive mutation on activity of the RNA transcriptase of vesicular stomatitis virus New Jersey. J Virol. 1979 Jun;30(3):692–700. doi: 10.1128/jvi.30.3.692-700.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Unger J. T., Reichmann M. E. RNA synthesis in temperature-sensitive mutants of vesicular stomatitis virus. J Virol. 1973 Sep;12(3):570–578. doi: 10.1128/jvi.12.3.570-578.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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