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Journal of Virology, January 2007, p. 613-619, Vol. 81, No. 2
0022-538X/07/$08.00+0     doi:10.1128/JVI.01668-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Assembly of Two Independent Populations of Flock House Virus Particles with Distinct RNA Packaging Characteristics in the Same Cell{triangledown} ,{dagger}

P. Arno Venter and Anette Schneemann*

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

Received 2 August 2006/ Accepted 25 October 2006

Flock House virus (FHV; Nodaviridae) is a positive-strand RNA virus that encapsidates a bipartite genome consisting of RNA1 and RNA2. We recently showed that specific recognition of these RNAs for packaging into progeny particles requires coat protein translated from replicating viral RNA. In the present study, we investigated whether the entire assembly pathway, i.e., the formation of the initial nucleating complex and the subsequent completion of the capsid, is restricted to the same pool of coat protein subunits. To test this, coat proteins carrying either FLAG or hemagglutinin epitopes were synthesized from replicating or nonreplicating RNA in the same cell, and the resulting particle population and its RNA packaging phenotype were analyzed. Results from immunoprecipitation analysis and ion-exchange chromatography showed that the differentially tagged proteins segregated into two distinct populations of virus particles with distinct RNA packaging phenotypes. Particles assembled from coat protein that was translated from replicating RNA contained the FHV genome, whereas particles assembled from coat protein that was translated from nonreplicating mRNA contained random cellular RNA. These data demonstrate that only coat proteins synthesized from replicating RNA partake in the assembly of virions that package the viral genome and that RNA replication, coat protein translation, and virion assembly are processes that are tightly coupled during the life cycle of FHV.

* Corresponding author. Mailing address: Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037. Phone: (858) 784-8643. Fax: (858) 784-7979. E-mail: aschneem{at}scripps.edu.

{triangledown} Published ahead of print on 1 November 2006.

{dagger} Manuscript 18190-MB from the Scripps Research Institute.

Journal of Virology, January 2007, p. 613-619, Vol. 81, No. 2
0022-538X/07/$08.00+0     doi:10.1128/JVI.01668-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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