Identification of an RNA-silencing suppressor in the genome of Grapevine virus A

J Gen Virol. 2006 Aug;87(Pt 8):2387-2395. doi: 10.1099/vir.0.81893-0.

Abstract

Higher plants use post-transcriptional gene silencing (PTGS), an RNA-degradation system, as a defence mechanism against viral infections. To counteract this, plant viruses encode and express PTGS suppressor proteins. Four of the five proteins encoded by the Grapevine virus A (GVA) genome were screened using a green fluorescent protein (GFP)-based transient expression assay, and the expression product of ORF5 (protein p10) was identified as a suppressor of silencing. ORF5 p10 suppressed local and systemic silencing induced by a transiently expressed single-stranded sense RNA. This protein was active towards both a transgene and exogenous GFP mRNAs. Ectopic expression of GVA-ORF5 by a Potato virus X vector enhanced symptom severity. The findings that p10 markedly reduces the levels of small interfering RNAs (siRNAs) and that the recombinant protein is able to bind single-stranded and double-stranded forms of siRNAs and microRNAs, suggest the existence of a potential mechanism of suppression based on RNA sequestering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Expression
  • Genes, Reporter
  • Genetic Vectors
  • Green Fluorescent Proteins / analysis
  • Nicotiana / virology
  • Plant Diseases / virology
  • Plant Viruses / genetics*
  • Plant Viruses / pathogenicity
  • Potexvirus / genetics
  • Protein Binding
  • RNA Interference*
  • RNA Viruses / genetics*
  • RNA Viruses / pathogenicity
  • RNA, Small Interfering / metabolism
  • Transfection
  • Viral Proteins / physiology*

Substances

  • RNA, Small Interfering
  • Viral Proteins
  • Green Fluorescent Proteins