This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Polo, J M Articles by Johnston, R E Search for Related Content PubMed PubMed Citation Articles by Polo, J M Articles by Johnston, R E

 Previous Article  |  Next Article 

J Virol. 1988 June; 62(6): 2124-2133

Molecular analysis of Sindbis virus pathogenesis in neonatal mice by using virus recombinants constructed in vitro.

Department of Microbiology, North Carolina State University, Raleigh 27695-7615.

ABSTRACT

Genetic loci affecting Sindbis virus pathogenesis in neonatal mice have been examined by using a full-length cDNA clone of the virus (Toto1101). The full-length cDNA is linked to a bacteriophage SP6 promoter to facilitate the synthesis of infectious RNA transcripts in vitro. Virus derived from Toto1101 showed reduced virulence (attenuation) in neonatal mice. Replacement of the E1 glycoprotein and 6K genes of Toto1101 with cloned E1 and 6K genes derived from a virulent Sindbis virus strain, AR339 (SB), resulted in a new construct, TR2000, that gave rise to virulent virus. Sequence determinations for the entire substituted regions of TR2000, Toto1101, and related virulent and attenuated strains identified three coding differences in E1 between Toto1101 and TR2000. These differences, individually or in combination, may be responsible for the attenuated phenotype. Previous studies in this laboratory identified another attenuating mutation at amino acid position 114 of the E2 glycoprotein (N.L. Davis, F.J. Fuller, W.G. Dougherty, R.A. Olmsted, and R.E. Johnston, Proc. Natl. Acad. Sci. USA 83:6771-6775, 1986). Substitution of Arg-114 in the mutant SB-RL for Ser-114 of SB appears to confer three distinguishing phenotypes: attenuation in neonatal mice, increased sensitivity to specific E2 monoclonal antibodies, and accelerated penetration of BHK cells. Replacement of TR2000 sequences containing the codon for amino acid 114 of E2 with corresponding fragments from cDNA clones of SB or SB-RL produced two strains of Sindbis virus (TR2100 and TR2200) which were isogenic except for the E2 114 codon (Ser and Arg, respectively). The three diagnostic phenotypes cosegregated according to the origin of the codon for amino acid 114 of E2, confirming the dramatic effect of this single amino acid substitution on these three phenotypes.

This article has been cited by other articles:

• Nagata, L. P., Hu, W.-G., Parker, M., Chau, D., Rayner, G. A., Schmaltz, F. L., Wong, J. P. (2006). Infectivity variation and genetic diversity among strains of Western equine encephalitis virus.. J. Gen. Virol. 87: 2353-2361 [Abstract] [Full Text]  
• Perri, S., Greer, C. E., Thudium, K., Doe, B., Legg, H., Liu, H., Romero, R. E., Tang, Z., Bin, Q., Dubensky, T. W. Jr., Vajdy, M., Otten, G. R., Polo, J. M. (2003). An Alphavirus Replicon Particle Chimera Derived from Venezuelan Equine Encephalitis and Sindbis Viruses Is a Potent Gene-Based Vaccine Delivery Vector. J. Virol. 77: 10394-10403 [Abstract] [Full Text]  
• Gardner, J. P., Frolov, I., Perri, S., Ji, Y., MacKichan, M. L., zur Megede, J., Chen, M., Belli, B. A., Driver, D. A., Sherrill, S., Greer, C. E., Otten, G. R., Barnett, S. W., Liu, M. A., Dubensky, T. W., Polo, J. M. (2000). Infection of Human Dendritic Cells by a Sindbis Virus Replicon Vector Is Determined by a Single Amino Acid Substitution in the E2 Glycoprotein. J. Virol. 74: 11849-11857 [Abstract] [Full Text]  
• Ryman, K. D., Klimstra, W. B., Nguyen, K. B., Biron, C. A., Johnston, R. E. (2000). Alpha/Beta Interferon Protects Adult Mice from Fatal Sindbis Virus Infection and Is an Important Determinant of Cell and Tissue Tropism. J. Virol. 74: 3366-3378 [Abstract] [Full Text]  
• Kobiler, D., Rice, C. M., Brodie, C., Shahar, A., Dubuisson, J., Halevy, M., Lustig, S. (1999). A Single Nucleotide Change in the 5' Noncoding Region of Sindbis Virus Confers Neurovirulence in Rats. J. Virol. 73: 10440-10446 [Abstract] [Full Text]