Nodavirus Rna Replication Protein a Induces Membrane Association of Genomic Rna.
From: Institute for Molecular Virology, University of Wisconsin-Madison, 1525 Linden Drive, Madison, WI 53706-1596, USA.
Journal of virology
- Publish Date: May 2007
- ISSN: 0022-538X
- Volume: 81
- Issue: 9
- Pages: 4633-44
- Medium: Print
- Language: English
- Citation (JAMA): Van Wynsberghe Priscilla M, Chen Hau-Ren, Ahlquist Paul, et al. Nodavirus Rna Replication Protein a Induces Membrane Association of Genomic Rna.. J. Virol. May 2007;81:4633-44
Abstract
Positive-strand RNA virus genome replication occurs in membrane-associated RNA replication complexes, whose assembly remains poorly understood. Here we show that prior to RNA replication, the multifunctional, transmembrane RNA replication protein A of the nodavirus flock house virus (FHV) recruits FHV genomic RNA1 to a membrane-associated state in both Drosophila melanogaster and Saccharomyces cerevisiae cells. Protein A has mitochondrial membrane-targeting, self-interaction, RNA-dependent RNA polymerase (RdRp), and RNA capping domains. In the absence of RdRp activity due to an active site mutation (A(D692E)), protein A stimulated RNA1 accumulation by increasing RNA1 stability. Protein A(D692E) stimulated RNA1 accumulation in wild-type cells and in xrn1(-) yeast defective in decapped RNA decay, showing that increased RNA1 stability was not due to protein A-mediated RNA1 recapping. Increased RNA1 stability was closely linked with protein A-induced membrane association of the stabilized RNA and was highly selective for RNA1. Substantial N- and C-proximal regions of protein A were dispensable for these activities. However, increased RNA1 accumulation was eliminated by deleting protein A amino acids (aa) 1 to 370 but was restored completely by adding back the transmembrane domain (aa 1 to 35) and partially by adding back peripheral membrane association sequences in aa 36 to 370. Moreover, although RNA polymerase activity was not required, even small deletions in or around the RdRp domain abolished increased RNA1 accumulation. These and other results show that prior to negative-strand RNA synthesis, multiple domains of mitochondrially targeted protein A cooperate to selectively recruit FHV genomic RNA to membranes where RNA replication complexes form.
Mesh Headings (Keywords): Animals, Blotting, Northern, Blotting, Western, Cells, Cultured, Drosophila melanogaster, Genomic Instability, Mutation, Nodaviridae, Protein Structure, Tertiary, RNA Replicase, RNA, Viral, Replication Protein A, Reverse Transcriptase Polymerase Chain Reaction, Saccharomyces cerevisiae, Virus Replication
Check for Full Text / PubMed Unique Identifier (PMID): 17301137
This abstract is part of PubMed, a service of the U.S. National Library of Medicine. PubMed includes more than 17 million citations from MEDLINE and other life science journals for biomedical articles. See Copyright and Disclaimers.
Linked medical terms appearing on this page are added by Healia to help readers find more information and are not part of the original PubMed document.
The data herein was last updated on July 8th, 2008 and may not reflect the most current and accurate data available from NLM.