RRC ID 69943
Author Hyodo K, Mine A, Taniguchi T, Kaido M, Mise K, Taniguchi H, Okuno T.
Title ADP ribosylation factor 1 plays an essential role in the replication of a plant RNA virus.
Journal J Virol
Abstract Eukaryotic positive-strand RNA viruses replicate using the membrane-bound replicase complexes, which contain multiple viral and host components. Virus infection induces the remodeling of intracellular membranes. Virus-induced membrane structures are thought to increase the local concentration of the components that are required for replication and provide a scaffold for tethering the replicase complexes. However, the mechanisms underlying virus-induced membrane remodeling are poorly understood. RNA replication of red clover necrotic mosaic virus (RCNMV), a positive-strand RNA plant virus, is associated with the endoplasmic reticulum (ER) membranes, and ER morphology is perturbed in RCNMV-infected cells. Here, we identified ADP ribosylation factor 1 (Arf1) in the affinity-purified RCNMV RNA-dependent RNA polymerase fraction. Arf1 is a highly conserved, ubiquitous, small GTPase that is implicated in the formation of the coat protein complex I (COPI) vesicles on Golgi membranes. Using in vitro pulldown and bimolecular fluorescence complementation analyses, we showed that Arf1 interacted with the viral p27 replication protein within the virus-induced large punctate structures of the ER membrane. We found that inhibition of the nucleotide exchange activity of Arf1 using the inhibitor brefeldin A (BFA) disrupted the assembly of the viral replicase complex and p27-mediated ER remodeling. We also showed that BFA treatment and the expression of dominant negative Arf1 mutants compromised RCNMV RNA replication in protoplasts. Interestingly, the expression of a dominant negative mutant of Sar1, a key regulator of the biogenesis of COPII vesicles at ER exit sites, also compromised RCNMV RNA replication. These results suggest that the replication of RCNMV depends on the host membrane traffic machinery.
Volume 87(1)
Pages 163-76
Published 2013-1-1
DOI 10.1128/JVI.02383-12
PII JVI.02383-12
PMID 23097452
PMC PMC3536388
MeSH ADP-Ribosylation Factor 1 / metabolism* Arabidopsis / virology* Centrifugation Endoplasmic Reticulum / virology Fluorescence Host-Pathogen Interactions* Protein Binding Protein Interaction Mapping Tobacco / virology* Tombusviridae / physiology* Viral Proteins / metabolism* Virus Replication*
IF 4.501
Arabidopsis / Cultured plant cells, genes rpc00001