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Journal of Virology, May 2005, p. 6239-6248, Vol. 79, No. 10
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.10.6239-6248.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Capsid Protein Synthesis from Replicating RNA Directs Specific Packaging of the Genome of a Multipartite, Positive-Strand RNA Virus{dagger}

P. Arno Venter, Neel K. Krishna,{ddagger} and Anette Schneemann*

Department of Molecular Biology, The Scripps Research Institute, La Jolla, California 92037

Received 12 October 2004/ Accepted 18 January 2005

Flock house virus (FHV) is a bipartite, positive-strand RNA insect virus that encapsidates its two genomic RNAs in a single virion. It provides a convenient model system for studying the principles underlying the copackaging of multipartite viral RNA genomes. In this study, we used a baculovirus expression system to determine if the uncoupling of viral protein synthesis from RNA replication affected the packaging of FHV RNAs. We found that neither RNA1 (which encodes the viral replicase) nor RNA2 (which encodes the capsid protein) were packaged efficiently when capsid protein was supplied in trans from nonreplicating RNA. However, capsid protein synthesized in cis from replicating RNA2 packaged RNA2 efficiently in the presence and absence of RNA1. These results demonstrated that capsid protein translation from replicating RNA2 is required for specific packaging of the FHV genome. This type of coupling between genome replication and translation and RNA packaging has not been observed previously. We hypothesize that RNA2 replication and translation must be spatially coordinated in FHV-infected cells to facilitate retrieval of the viral RNAs for encapsidation by newly synthesized capsid protein. Spatial coordination of RNA and capsid protein synthesis may be key to specific genome packaging and assembly in other RNA viruses.

* Corresponding author. Mailing address: Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037. Phone: (858) 784-8643. Fax: (858) 784-8660. E-mail: aschneem{at}scripps.edu.

{dagger} Manuscript 16850-MB from the Scripps Research Institute.

{ddagger} Present address: Department of Pediatrics and the Center for Pediatric Research, Eastern Virginia Medical School and Children's Hospital of the King's Daughters, Norfolk, VA 23510.

Journal of Virology, May 2005, p. 6239-6248, Vol. 79, No. 10
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.10.6239-6248.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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