On the nature of in vivo requirements for rde-4 in RNAi and developmental pathways in C. elegans

RNA Biol. 2011 May-Jun;8(3):458-67. doi: 10.4161/rna.8.3.14657. Epub 2011 May 1.

Abstract

C. elegans RDE-4 is a double-stranded RNA binding protein that has been shown to play a key role in response to foreign double-stranded RNA (dsRNA). We have used diverse tools for analysis of gene function to characterize the domain and organismal foci of RDE-4 action in C. elegans. First, we examined the focus of activity within the RDE-4 protein, by testing a series of RDE-4 deletion constructs for their ability to support dsRNA-triggered gene silencing. These assays indicated a molecular requirement for a linker region and the second dsRNA-binding domain of RDE-4, with ancillary contributions to function from the C and N terminal domains. Second, we used mosaic analysis to explore the cellular focus of action of RDE-4. These experiments indicated an ability of RDE-4 to function non-autonomously in foreign RNA responses. Third, we used growth under stressful conditions to search for evidence of an organismal focus of action for RDE-4 distinct from its role in response to foreign dsRNA. Propagation at high temperatures exposed a conditional requirement for RDE-4 for optimal growth and fertility, indicating at least under these conditions that RDE-4 can serve an essential role in C. elegans.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Binding Sites
  • Caenorhabditis elegans / embryology
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans Proteins / genetics*
  • Caenorhabditis elegans Proteins / metabolism*
  • Gene Silencing
  • RNA Interference*
  • RNA, Double-Stranded / metabolism
  • RNA, Helminth / metabolism
  • RNA-Binding Proteins / genetics*
  • RNA-Binding Proteins / metabolism*
  • Temperature

Substances

  • Caenorhabditis elegans Proteins
  • RDE-4 protein, C elegans
  • RNA, Double-Stranded
  • RNA, Helminth
  • RNA-Binding Proteins