This Article Full Text 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 Sawicki, D. L. Articles by Sawicki, S. G. Search for Related Content PubMed PubMed Citation Articles by Sawicki, D. L. Articles by Sawicki, S. G.

Alphavirus Minus-Strand Synthesis and Persistence in Mouse Embryo Fibroblasts Derived from Mice Lacking RNase L and Protein Kinase R

Department of Microbiology and Immunology, Medical College of Ohio, Toledo, Ohio 43614,¹ Lerner Research Institute, The Cleveland Clinic Foundation, Cleveland, Ohio 44195²

Received 16 July 2002/ Accepted 24 October 2002

We report our studies to probe the possible role of the host response to double-stranded RNA in cessation of alphavirus minus-strand synthesis. Mouse embryo fibroblasts (MEF) from Mx1-deficient mice that also lack either the protein kinase R (PKR) or the latent RNase L or both PKR and RNase L were screened. In RNase L-deficient but not wild-type or PKR-deficient MEF, there was continuous synthesis of minus-strand templates and the formation of new replication complexes producing viral plus strands. Inhibiting translation caused minus-strand synthesis to stop and a loss of transcription activity of the mature replication complexes. This turnover of replication complexes that were stable in cells containing RNase L suggested that RNase L plays some role, albeit possibly indirect, in the formation of stable replication complexes during alphavirus infection. In addition, confluent monolayers of RNase L-deficient murine cells readily established persistent infections and were not killed. This phenotype is contrary to what has been observed for infection in vertebrate cells with a presumably functional RNase L gene and more resembled alphavirus replication in Aedes mosquito cells, in which the activity of replication complexes making plus stands was also found to decay with inhibition of translation.

This article has been cited by other articles:

• Mayuri, , Geders, T. W., Smith, J. L., Kuhn, R. J. (2008). Role for Conserved Residues of Sindbis Virus Nonstructural Protein 2 Methyltransferase-Like Domain in Regulation of Minus-Strand Synthesis and Development of Cytopathic Infection. J. Virol. 82: 7284-7297 [Abstract] [Full Text]  
• Gorchakov, R., Frolova, E., Sawicki, S., Atasheva, S., Sawicki, D., Frolov, I. (2008). A New Role for ns Polyprotein Cleavage in Sindbis Virus Replication. J. Virol. 82: 6218-6231 [Abstract] [Full Text]  
• Silverman, R. H. (2007). Viral Encounters with 2',5'-Oligoadenylate Synthetase and RNase L during the Interferon Antiviral Response. J. Virol. 81: 12720-12729 [Full Text]  
• Samuel, M. A., Whitby, K., Keller, B. C., Marri, A., Barchet, W., Williams, B. R. G., Silverman, R. H., Gale, M. Jr., Diamond, M. S. (2006). PKR and RNase L Contribute to Protection against Lethal West Nile Virus Infection by Controlling Early Viral Spread in the Periphery and Replication in Neurons. J. Virol. 80: 7009-7019 [Abstract] [Full Text]  
• Sawicki, D. L., Perri, S., Polo, J. M., Sawicki, S. G. (2006). Role for nsP2 Proteins in the Cessation of Alphavirus Minus-Strand Synthesis by Host Cells. J. Virol. 80: 360-371 [Abstract] [Full Text]  
• Ryman, K. D., Meier, K. C., Nangle, E. M., Ragsdale, S. L., Korneeva, N. L., Rhoads, R. E., MacDonald, M. R., Klimstra, W. B. (2005). Sindbis Virus Translation Is Inhibited by a PKR/RNase L-Independent Effector Induced by Alpha/Beta Interferon Priming of Dendritic Cells. J. Virol. 79: 1487-1499 [Abstract] [Full Text]  
• De, I., Fata-Hartley, C., Sawicki, S. G., Sawicki, D. L. (2003). Functional Analysis of nsP3 Phosphoprotein Mutants of Sindbis Virus. J. Virol. 77: 13106-13116 [Abstract] [Full Text]