Abstract
Three types of conditional lethal mutant were isolated from wild-type vesicular stomatitis virus, New Jersey serotype, after mutagenization by 5-fluorouracil: (i) conventional temperature-sensitive (ts) mutants, which form plaques at 31 C but not at 39 C; (ii) conventional host range mutants (hr CE), which grow in BHK but not in secondary chicken embryo cells; and (iii) temperature-dependent host range mutants (td CE), which form plaques both at 31 and 39 C on BHK cells but only at 31 C on chicken embryo cells. To determine whether the mutation in hr CE and td CE mutants affected the virion-associated RNA transcriptase, this enzyme was assayed in vitro at 31 and 39 C, and the results were compared with those obtained for the wild-type virus. The RNA trascriptase activity of hr CE mutants did not appear to be affected by the mutation. The td CE mutants fall into two classes: those that synthesized RNA at 39 C similar to the wild-type virus and those that did not. One mutant of the latter category, td CE 3, had heat-sensitive transcriptase regardless of whether it was grown in BHK or chicken embryo cells. A revertant to the wild-type phenotype isolated from this mutant had regained the ability to synthesize RNA at 39 C. These results strongly suggest that a polypeptide that is either the transcriptase itself or part of the transcriptase complex was made temperature sensitive by the mutation in the second class of td CE mutants. The inhibition of the transcriptase activity of the mutant td CE 3 was fully reversible by lowering the temperature of incubation from 39 to 31 C, and both inhibition and reactivation appeared to be instantaneous.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- Pringle C. R. Genetic characteristics of conditional lethal mutants of vesicular stomatitis virus induced by 5-fluorouracil, 5-azacytidine, and ethyl methane sulfonate. J Virol. 1970 May;5(5):559–567. doi: 10.1128/jvi.5.5.559-567.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rettenmier C. W., Dumont R., Baltimore D. Screening procedure for complementation-dependent mutants of vesicular stomatitis virus. J Virol. 1975 Jan;15(1):41–49. doi: 10.1128/jvi.15.1.41-49.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simpson R. W., Obijeski J. F. Conditional lethal mutants of vesicular stomatitis virus. I. Phenotypic characterization of single and double mutants exhibiting host restriction and temperature sensitivity. Virology. 1974 Feb;57(2):357–368. doi: 10.1016/0042-6822(74)90175-5. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]