Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1991 Nov;65(11):6358–6361. doi: 10.1128/jvi.65.11.6358-6361.1991

Mutational analysis of a virulence locus in the E2 glycoprotein gene of Sindbis virus.

J M Polo 1, R E Johnston 1
PMCID: PMC250357  PMID: 1656101

Abstract

The substitution of arginine for serine at position 114 of glycoprotein E2 in several biological and recombinant Sindbis virus mutants was shown previously to attenuate the virus for neonatal mice and also to accelerate virus penetration into BHK cells. To further examine the genetically linked effects on both virus penetration into cultured cells and pathogenesis in vivo, mutants containing each of 16 different amino acid coding changes at this position were generated by site-directed mutagenesis of a full-length cDNA clone of the Sindbis virus genome. Viable virus was recovered following transfection of RNA transcripts from 14 of the clones. Phenotypic analysis of these virus mutants revealed that specific amino acid residues affected either the pathogenesis or penetration phenotype independently or both phenotypes simultaneously. Thus, both the position of a mutation within the E2 sequence and the particular amino acid encoded at that position are important determinants of the mutant phenotypes.

Full text

PDF
6358

Selected References

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

  1. Baric R. S., Trent D. W., Johnston R. E. A Sindbis virus variant with a cell-determined latent period. Virology. 1981 Apr 15;110(1):237–242. doi: 10.1016/0042-6822(81)90029-5. [DOI] [PubMed] [Google Scholar]
  2. Bose H. R., Sagik B. P. The virus envelope in cell attachment. J Gen Virol. 1970 Nov;9(2):159–161. doi: 10.1099/0022-1317-9-2-159. [DOI] [PubMed] [Google Scholar]
  3. Davis N. L., Fuller F. J., Dougherty W. G., Olmsted R. A., Johnston R. E. A single nucleotide change in the E2 glycoprotein gene of Sindbis virus affects penetration rate in cell culture and virulence in neonatal mice. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6771–6775. doi: 10.1073/pnas.83.18.6771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Flynn D. C., Meyer W. J., Mackenzie J. M., Jr, Johnston R. E. A conformational change in Sindbis virus glycoproteins E1 and E2 is detected at the plasma membrane as a consequence of early virus-cell interaction. J Virol. 1990 Aug;64(8):3643–3653. doi: 10.1128/jvi.64.8.3643-3653.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Flynn D. C., Olmsted R. A., Mackenzie J. M., Jr, Johnston R. E. Antibody-mediated activation of Sindbis virus. Virology. 1988 Sep;166(1):82–90. doi: 10.1016/0042-6822(88)90149-3. [DOI] [PubMed] [Google Scholar]
  6. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  7. Lustig S., Jackson A. C., Hahn C. S., Griffin D. E., Strauss E. G., Strauss J. H. Molecular basis of Sindbis virus neurovirulence in mice. J Virol. 1988 Jul;62(7):2329–2336. doi: 10.1128/jvi.62.7.2329-2336.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Olmsted R. A., Baric R. S., Sawyer B. A., Johnston R. E. Sindbis virus mutants selected for rapid growth in cell culture display attenuated virulence in animals. Science. 1984 Jul 27;225(4660):424–427. doi: 10.1126/science.6204381. [DOI] [PubMed] [Google Scholar]
  9. Olmsted R. A., Meyer W. J., Johnston R. E. Characterization of Sindbis virus epitopes important for penetration in cell culture and pathogenesis in animals. Virology. 1986 Jan 30;148(2):245–254. doi: 10.1016/0042-6822(86)90322-3. [DOI] [PubMed] [Google Scholar]
  10. Pence D. F., Davis N. L., Johnston R. E. Antigenic and genetic characterization of Sindbis virus monoclonal antibody escape mutants which define a pathogenesis domain on glycoprotein E2. Virology. 1990 Mar;175(1):41–49. doi: 10.1016/0042-6822(90)90184-s. [DOI] [PubMed] [Google Scholar]
  11. Polo J. M., Davis N. L., Rice C. M., Huang H. V., Johnston R. E. Molecular analysis of Sindbis virus pathogenesis in neonatal mice by using virus recombinants constructed in vitro. J Virol. 1988 Jun;62(6):2124–2133. doi: 10.1128/jvi.62.6.2124-2133.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Polo J. M., Johnston R. E. Attenuating mutations in glycoproteins E1 and E2 of Sindbis virus produce a highly attenuated strain when combined in vitro. J Virol. 1990 Sep;64(9):4438–4444. doi: 10.1128/jvi.64.9.4438-4444.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Rice C. M., Levis R., Strauss J. H., Huang H. V. Production of infectious RNA transcripts from Sindbis virus cDNA clones: mapping of lethal mutations, rescue of a temperature-sensitive marker, and in vitro mutagenesis to generate defined mutants. J Virol. 1987 Dec;61(12):3809–3819. doi: 10.1128/jvi.61.12.3809-3819.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Russell D. L., Dalrymple J. M., Johnston R. E. Sindbis virus mutations which coordinately affect glycoprotein processing, penetration, and virulence in mice. J Virol. 1989 Apr;63(4):1619–1629. doi: 10.1128/jvi.63.4.1619-1629.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Stanley J., Cooper S. J., Griffin D. E. Alphavirus neurovirulence: monoclonal antibodies discriminating wild-type from neuroadapted Sindbis virus. J Virol. 1985 Oct;56(1):110–119. doi: 10.1128/jvi.56.1.110-119.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Zimmern D., Kaesberg P. 3'-terminal nucleotide sequence of encephalomyocarditis virus RNA determined by reverse transcriptase and chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4257–4261. doi: 10.1073/pnas.75.9.4257. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES