RRC ID 46394
Author LeGendre JB, Campbell ZT, Kroll-Conner P, Anderson P, Kimble J, Wickens M.
Title RNA targets and specificity of Staufen, a double-stranded RNA-binding protein in Caenorhabditis elegans.
Journal J Biol Chem
Abstract The Staufen family consists of proteins that possess double-stranded RNA-binding domains (dsRBDs). Staufen proteins of Drosophila and mammals regulate mRNA localization, translation, and decay. We report analysis of Staufen in Caenorhabditis elegans, which we have designated STAU-1. We focus on its biochemical properties, mRNA targets, and possible role in RNAi. We show that STAU-1 is expressed as mRNA and protein at all stages of C. elegans development. The wild-type, full-length protein, purified from bacteria, binds duplex RNA with high affinity in vitro. Purified, mutant proteins lacking single dsRBDs still bind RNA efficiently, demonstrating that no single domain is required for binding to duplex RNA (although dsRBD2 could not be tested). STAU-1 mRNA targets were identified via immunoprecipitation with specific anti-STAU-1 antibodies, followed by microarray analysis (RIP-Chip). These studies define a set of 418 likely STAU-1 mRNA targets. Finally, we demonstrate that stau-1 mutants enhance exogenous RNAi and that stau-1;eri-1 double mutants exhibit sterility and synthetic germ line defects.
Volume 288(4)
Pages 2532-45
Published 2013-1-25
DOI 10.1074/jbc.M112.397349
PII M112.397349
PMID 23195953
PMC PMC3554921
MeSH 3' Untranslated Regions Animals Biochemistry / methods Caenorhabditis elegans Caenorhabditis elegans Proteins / chemistry* Caenorhabditis elegans Proteins / physiology DNA, Complementary / metabolism Models, Biological Mutation Oligonucleotide Array Sequence Analysis Protein Binding RNA Interference RNA, Messenger / metabolism RNA-Binding Proteins / chemistry* RNA-Binding Proteins / physiology Transgenes Two-Hybrid System Techniques
IF 4.106
Times Cited 35
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
C.elegans tm2266