Analysis of siRNA-suppressor of gene silencing interactions

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

RNA silencing is an evolutionarily conserved system that functions as an antiviral mechanism in higher plants and animals. To counteract RNA silencing, viruses evolved silencing suppressors that interfere with siRNA guided RNA silencing pathway. We used the heterologous Drosophila in vitro embryo RNA to analyze the molecular mechanism of suppression of silencing suppressors. We found that different silencing suppressors inhibit the RNA silencing via binding to siRNAs. None of the suppressors affected the activity of preassembled RISC complexes. In contrast, suppressors uniformly inhibited the siRNA-initiated RISC assembly pathway by preventing RNA silencing initiator complex formation. Here, we provide the protocol for the detailed analysis of p19 silencing suppressors of tombusviruses in the heterologous Drosophila in vitro system.

Original languageEnglish
Title of host publicationPlant Virology Protocols
Subtitle of host publicationFrom Viral Sequence to Protein Function
PublisherHumana Press
Pages331-337
Number of pages7
ISBN (Print)9781588298270
DOIs
Publication statusPublished - Jan 1 2008

Publication series

NameMethods in Molecular Biology
Volume451
ISSN (Print)1064-3745

Keywords

  • In vitro Drosophila RNA silencing system
  • P19
  • SiRNA
  • SiRNA binding
  • Silencing suppressor

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Lakatos, L., & Burgyán, J. (2008). Analysis of siRNA-suppressor of gene silencing interactions. In Plant Virology Protocols: From Viral Sequence to Protein Function (pp. 331-337). (Methods in Molecular Biology; Vol. 451). Humana Press. https://doi.org/10.1007/978-1-59745-102-4_23