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 Chen, J. Articles by Ahlquist, P. Search for Related Content PubMed PubMed Citation Articles by Chen, J. Articles by Ahlquist, P.

An Alternate Pathway for Recruiting Template RNA to the Brome Mosaic Virus RNA Replication Complex

Institute for Molecular Virology,¹ Howard Hughes Medical Institute, University of Wisconsin, Madison, Wisconsin 53706²

Received 14 August 2002/ Accepted 12 November 2002

The multidomain RNA replication protein 1a of brome mosaic virus (BMV), a positive-strand RNA virus in the alphavirus-like superfamily, plays key roles in assembly and function of the viral RNA replication complex. 1a, which encodes RNA capping and helicase-like domains, localizes to endoplasmic reticulum membranes, recruits BMV 2a polymerase and viral RNA templates, and forms membrane-bound, capsid-like spherules in which RNA replication occurs. cis-acting signals necessary and sufficient for RNA recruitment by 1a have been mapped in BMV genomic RNA2 and RNA3. Both signals comprise an extended stem-loop whose apex matches the conserved sequence and structure of the TC stem-loop in tRNAs (box B). Mutations show that this box B motif is crucial to 1a responsiveness of wild-type RNA2 and RNA3. We report here that, unexpectedly, some chimeric mRNAs expressing the 2a polymerase open reading frame from RNA2 were recruited by 1a to the replication complex and served as templates for negative-strand RNA synthesis, despite lacking the normally essential, box B-containing 5' signal. Further studies showed that this template recruitment required high-efficiency translation of the RNA templates. Moreover, multiple small frameshifting insertion or deletion mutations throughout the N-terminal region of the open reading frame inhibited this template recruitment, while an in-frame insertion did not. Providing 2a in trans did not restore template recruitment of RNAs with frameshift mutations. Only those deletions in the N-terminal region of 2a that abolished 2a interaction with 1a abolished template recruitment of the RNA. These and other results indicate that this alternate pathway for 1a-dependent RNA recruitment involves 1a interaction with the translating mRNA via the 1a-interactive N-terminal region of the nascent 2a polypeptide. Interaction with nascent 2a also may be involved in 1a recruitment of 2a polymerase to membranes.

This article has been cited by other articles:

• Yi, G., Kao, C. (2008). cis- and trans-Acting Functions of Brome Mosaic Virus Protein 1a in Genomic RNA1 Replication. J. Virol. 82: 3045-3053 [Abstract] [Full Text]  
• Taylor, M. P., Kirkegaard, K. (2007). Modification of Cellular Autophagy Protein LC3 by Poliovirus. J. Virol. 81: 12543-12553 [Abstract] [Full Text]  
• Zhu, J., Gopinath, K., Murali, A., Yi, G., Hayward, S. D., Zhu, H., Kao, C. (2007). RNA-binding proteins that inhibit RNA virus infection. Proc. Natl. Acad. Sci. USA 104: 3129-3134 [Abstract] [Full Text]  
• Yi, G., Gopinath, K., Kao, C. C. (2007). Selective Repression of Translation by the Brome Mosaic Virus 1a RNA Replication Protein. J. Virol. 81: 1601-1609 [Abstract] [Full Text]  
• Pijlman, G. P., Kondratieva, N., Khromykh, A. A. (2006). Translation of the Flavivirus Kunjin NS3 Gene in cis but Not Its RNA Sequence or Secondary Structure Is Essential for Efficient RNA Packaging. J. Virol. 80: 11255-11264 [Abstract] [Full Text]  
• Osman, T. A. M., Coutts, R. H. A., Buck, K. W. (2006). In Vitro Synthesis of Minus-Strand RNA by an Isolated Cereal Yellow Dwarf Virus RNA-Dependent RNA Polymerase Requires VPg and a Stem-Loop Structure at the 3' End of the Virus RNA. J. Virol. 80: 10743-10751 [Abstract] [Full Text]  
• Egger, D., Bienz, K. (2005). Intracellular location and translocation of silent and active poliovirus replication complexes. J. Gen. Virol. 86: 707-718 [Abstract] [Full Text]  
• Grdzelishvili, V. Z., Garcia-Ruiz, H., Watanabe, T., Ahlquist, P. (2005). Mutual Interference between Genomic RNA Replication and Subgenomic mRNA Transcription in Brome Mosaic Virus. J. Virol. 79: 1438-1451 [Abstract] [Full Text]  
• Choi, S.-K., Hema, M., Gopinath, K., Santos, J., Kao, C. (2004). Replicase-Binding Sites on Plus- and Minus-Strand Brome Mosaic Virus RNAs and Their Roles in RNA Replication in Plant Cells. J. Virol. 78: 13420-13429 [Abstract] [Full Text]  
• Miyanari, Y., Hijikata, M., Yamaji, M., Hosaka, M., Takahashi, H., Shimotohno, K. (2003). Hepatitis C Virus Non-structural Proteins in the Probable Membranous Compartment Function in Viral Genome Replication. J. Biol. Chem. 278: 50301-50308 [Abstract] [Full Text]  
• Vlot, A. C., Laros, S. M., Bol, J. F. (2003). Coordinate Replication of Alfalfa Mosaic Virus RNAs 1 and 2 Involves cis- and trans-Acting Functions of the Encoded Helicase-Like and Polymerase-Like Domains. J. Virol. 77: 10790-10798 [Abstract] [Full Text]