Rice, Charles M. and Franke, Christine A. and Strauss, James H. and Hruby, Dennis E. (1985) Expression of Sindbis virus structural proteins via recombinant vaccinia virus: synthesis, processing, and incorporation into mature Sindbis virions. Journal of Virology, 56 (1). pp. 227-239. ISSN 0022-538X. PMCID PMC252510. http://resolver.caltech.edu/CaltechAUTHORS:RICjvir85
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We have obtained a vaccinia virus recombinant which contains a complete cDNA copy of the 26S RNA of Sindbis virus within the thymidine kinase gene of the vaccinia virus genome. This recombinant constitutively transcribed the Sindbis sequences throughout the infectious cycle, reflecting the dual early-late vaccinia promoter used in this construction. The Sindbis-derived transcripts were translationally active, giving rise to both precursor and mature structural proteins of Sindbis virus, including the capsid protein (C), the precursor of glycoprotein E2 (PE2), and the two mature envelope glycoproteins (E1 and E2). These are the same products translated from the 26S mRNA during Sindbis infection, and thus these proteins were apparently cleaved, glycosylated, and transported in a manner analogous to that seen during authentic Sindbis infections. By using epitope-specific antibodies, it was possible to demonstrate that recombinant-derived proteins were incorporated into Sindbis virions during coinfections with monoclonal antibody-resistant Sindbis variants. These results suggest that all the information necessary to specify the proper biogenesis of Sindbis virus structural proteins resides within the 26S sequences and that vaccinia may provide an appropriate system for using DNA molecular genetic manipulations to unravel a variety of questions pertinent to RNA virus replication.
|Additional Information:||Copyright © 1985 by the American Society for Microbiology. Received 25 April 1985/Accepted 23 June 1985 We thank A. L. Schmaljohn and J. Dalrymple for providing epitope-specific anti-Sindbis monoclonal antibodies and resistant variants, B. Moss for providing pGS20, and Elias Lazarides and Seymour Benzer and the members of their laboratories for help with the indirect immunofluorescence. This research was supported by funds from the Murdoch Trust Foundation (to D.E.H.), by grants PCM-8316390 (to D.E.H.) and DMB 8316856 (to J.H.S.) from the National Science Foundation, by Public Health Service grants AI 10793 (to J.H.S.) and Al 20612 (to J.H.S.) from the National Institutes of Health, and by Biomedical Research Support grant 557 RR 07003.|
|PubMed Central ID:||PMC252510|
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|Deposited By:||Archive Administrator|
|Deposited On:||29 Nov 2006|
|Last Modified:||15 Dec 2015 22:43|
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