Interactions between p27 and p88 replicase proteins of Red clover necrotic mosaic virus play an essential role in viral RNA replication and suppression of RNA silencing via the 480-kDa viral replicase complex assembly

Akira Mine, Kiwamu Hyodo, Atsushi Takeda, Masanori Kaido, Kazuyuki Mise, Tetsuro Okuno

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Red clover necrotic mosaic virus (RCNMV), a positive-sense RNA virus with a bipartite genome, encodes p27 and p88 replicase proteins that are required for viral RNA replication and suppression of RNA silencing. In this study, we indentified domains in p27 and p88 responsible for their protein-protein interactions using in vitro pull-down assays with the purified recombinant proteins. Coimmunoprecipitation analysis in combination with blue-native polyacrylamide gel electrophoresis using mutated p27 proteins showed that both p27-p27 and p27-p88 interactions are essential for the formation of the 480-kDa complex, which has RCNMV-specific RNA-dependent RNA polymerase activity. Furthermore, we found a good correlation between the accumulated levels of the 480-kDa complex and replication levels and the suppression of RNA silencing activity. Our results indicate that interactions between RCNMV replicase proteins play an essential role in viral RNA replication and in suppressing RNA silencing via the 480-kDa replicase complex assembly.

Original languageEnglish
Pages (from-to)213-224
Number of pages12
JournalVirology
Volume407
Issue number2
DOIs
Publication statusPublished - Nov 25 2010
Externally publishedYes

Keywords

  • Blue-native PAGE
  • Coimmunoprecipitation analysis
  • Dianthovirus
  • In vitro translation and RNA synthesis
  • Positive-sense RNA virus
  • Protein-protein interaction
  • RNA silencing suppression
  • RNA-dependent RNA polymerase
  • Tombusviridae
  • Viral RNA replication
  • Viral replicase complex

ASJC Scopus subject areas

  • Virology

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