title: RNA targets and specificity of Staufen, a double-stranded RNA-binding protein in Caenorhabditis elegans.
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.
authors.affiliation: Graduate Program in Cellular and Molecular Biology, University of Wisconsin, Madison, Wisconsin 53706, USA.
authors.forename: Jacqueline Baca
authors.initials: JB
authors.lastname: LeGendre
authors.forename: Zachary T
authors.initials: ZT
authors.lastname: Campbell
authors.forename: Peggy
authors.initials: P
authors.lastname: Kroll-Conner
authors.forename: Phil
authors.initials: P
authors.lastname: Anderson
authors.forename: Judith
authors.initials: J
authors.lastname: Kimble
authors.forename: Marvin
authors.initials: M
authors.lastname: Wickens
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D001671:Biochemistry / Q000379:methods; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry* / Q000502:physiology; D018076:DNA, Complementary / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D011485:Protein Binding; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000502:physiology; D019076:Transgenes; D020798:Two-Hybrid System Techniques
chemical_list: D020413:3' Untranslated Regions; D029742:Caenorhabditis elegans Proteins; D018076:DNA, Complementary; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C579338:stau-1 proteins, C elegans