ADP ribosylation factor 1 plays an essential role in the replication of a plant RNA virus

Kiwamu Hyoudou, Akira Mine, Takako Taniguchi, Masanori Kaido, Kazuyuki Mise, Hisaaki Taniguchi, Tetsuro Okuno

Research output: Contribution to journalArticle

41 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)163-176
Number of pages14
JournalJournal of Virology
Volume87
Issue number1
DOIs
Publication statusPublished - Jan 2013
Externally publishedYes

Fingerprint

Red clover necrotic mosaic virus
ADP-Ribosylation Factor 1
Plant RNA
Tombusviridae
Plant Viruses
RNA Viruses
endoplasmic reticulum
Endoplasmic Reticulum
Membranes
brefeldin A
viruses
Brefeldin A
RNA
Virus Replication
Viruses
RNA-directed RNA polymerase
Coat Protein Complex I
mutants
RNA Replicase
Virus Assembly

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

ADP ribosylation factor 1 plays an essential role in the replication of a plant RNA virus. / Hyoudou, Kiwamu; Mine, Akira; Taniguchi, Takako; Kaido, Masanori; Mise, Kazuyuki; Taniguchi, Hisaaki; Okuno, Tetsuro.

In: Journal of Virology, Vol. 87, No. 1, 01.2013, p. 163-176.

Research output: Contribution to journalArticle

Hyoudou, K, Mine, A, Taniguchi, T, Kaido, M, Mise, K, Taniguchi, H & Okuno, T 2013, 'ADP ribosylation factor 1 plays an essential role in the replication of a plant RNA virus', Journal of Virology, vol. 87, no. 1, pp. 163-176. https://doi.org/10.1128/JVI.02383-12
Hyoudou, Kiwamu ; Mine, Akira ; Taniguchi, Takako ; Kaido, Masanori ; Mise, Kazuyuki ; Taniguchi, Hisaaki ; Okuno, Tetsuro. / ADP ribosylation factor 1 plays an essential role in the replication of a plant RNA virus. In: Journal of Virology. 2013 ; Vol. 87, No. 1. pp. 163-176.
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