PMID: 9658098Jul 11, 1998Paper

Template-dependent initiation of Sindbis virus RNA replication in vitro

Journal of Virology
J A LemmCharles M Rice

Abstract

Recent insights into the early events in Sindbis virus RNA replication suggest a requirement for either the P123 or P23 polyprotein, as well as mature nsP4, the RNA-dependent RNA polymerase, for initiation of minus-strand RNA synthesis. Based on this observation, we have succeeded in reconstituting an in vitro system for template-dependent initiation of SIN RNA replication. Extracts were isolated from cells infected with vaccinia virus recombinants expressing various SIN proteins and assayed by the addition of exogenous template RNAs. Extracts from cells expressing P123C>S, a protease-defective P123 polyprotein, and nsP4 synthesized a genome-length minus-sense RNA product. Replicase activity was dependent upon addition of exogenous RNA and was specific for alphavirus plus-strand RNA templates. RNA synthesis was also obtained by coexpression of nsP1, P23C>S, and nsP4. However, extracts from cells expressing nsP4 and P123, a cleavage-competent P123 polyprotein, had much less replicase activity. In addition, a P123 polyprotein containing a mutation in the nsP2 protease which increased the efficiency of processing exhibited very little, if any, replicase activity. These results provide further evidence that processing of the polypr...Continue Reading

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Citations

Aug 15, 2003·The Journal of Biological Chemistry·Lidia VasiljevaTero Ahola
Dec 3, 2003·Journal of Virology·Indra DéDorothea L Sawicki
Aug 7, 2002·Journal of Virology·Cori L FataDorothea L Sawicki
Jun 16, 2004·Proceedings of the National Academy of Sciences of the United States of America·Mei-Ling Li, Victor Stollar
Aug 31, 1999·The Journal of General Virology·E ten DamM D Ryan
Mar 31, 2006·Journal of Virology·Elena FrolovaIlya Frolov
May 29, 2004·The Journal of General Virology·Hai-He Wang, Sek-Man Wong
Jun 12, 2009·The Journal of General Virology·Eunhye Park, Diane E Griffin
Dec 1, 2001·Expert Opinion on Biological Therapy·S Schlesinger
Sep 5, 2002·Reviews in Medical Virology·Jonathan O RaynerKurt I Kamrud
Jul 27, 2007·Journal of Virology·Stephanie A Montgomery, Robert E Johnston
Dec 15, 2005·Journal of Virology·Dorothea L SawickiStanley G Sawicki
Jan 21, 2011·Journal of Virology·Jonathan C RuppRichard W Hardy
Mar 30, 2005·Journal of Virology·Richard W Hardy, Charles M Rice
Sep 3, 2009·Future Microbiology·Joyce JoseRichard J Kuhn
Jul 30, 2015·The Journal of General Virology·Jonathan C RuppRichard W Hardy
Mar 11, 2006·The Journal of General Virology·Sareen E GalbraithGregory J Atkins
Feb 10, 2018·Viruses·Farhana Abu Bakar, Lisa F P Ng
Aug 11, 2018·Frontiers in Molecular Biosciences·Anna K BlakneyRobin J Shattock

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