title: On the nature of in vivo requirements for rde-4 in RNAi and developmental pathways in C. elegans.
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.
authors.affiliation: Department of Pathology, Stanford University, Stanford, CA, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Blanchard
authors.forename: Poornima
authors.initials: P
authors.lastname: Parameswaran
authors.forename: Javier
authors.initials: J
authors.lastname: Lopez-Molina
authors.forename: Jonathan
authors.initials: J
authors.lastname: Gent
authors.forename: Jamie Fleenor
authors.initials: JF
authors.lastname: Saynuk
authors.forename: Andrew
authors.initials: A
authors.lastname: Fire
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001665:Binding Sites; D017173:Caenorhabditis elegans / Q000196:embryology / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics* / Q000378:metabolism*; D020868:Gene Silencing; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000378:metabolism; D018510:RNA, Helminth / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D013696:Temperature
chemical_list: D029742:Caenorhabditis elegans Proteins; C461645:RDE-4 protein, C elegans; D012330:RNA, Double-Stranded; D018510:RNA, Helminth; D016601:RNA-Binding Proteins