MUT-14 and SMUT-1 DEAD Box RNA Helicases Have Overlapping Roles in Germline RNAi and Endogenous siRNA Formation
| Autor: | Josien C. van Wolfswinkel, Peter C. Breen, Martin A. Newman, Brooke E Montgomery, Elke F. Roovers, René F. Ketting, Toshiro K. Ohsumi, Young-Soo Rim, Carolyn M. Phillips, Gary Ruvkun, Taiowa A. Montgomery |
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| Rok vydání: | 2014 |
| Předmět: |
Small interfering RNA
congenital hereditary and neonatal diseases and abnormalities DEAD box Trans-acting siRNA Fluoroimmunoassay Molecular Sequence Data Saccharomyces cerevisiae Biology Real-Time Polymerase Chain Reaction General Biochemistry Genetics and Molecular Biology Article DEAD-box RNA Helicases 03 medical and health sciences 0302 clinical medicine Mutant protein RNA interference Animals Immunoprecipitation RNA Small Interfering Caenorhabditis elegans Caenorhabditis elegans Proteins Gene 030304 developmental biology 0303 health sciences Base Sequence Agricultural and Biological Sciences(all) Biochemistry Genetics and Molecular Biology(all) Sequence Analysis DNA Molecular biology RNA Helicase A RNA silencing Germ Cells RNA Interference General Agricultural and Biological Sciences Sequence Alignment 030217 neurology & neurosurgery |
| Zdroj: | Current Biology |
| ISSN: | 0960-9822 |
| DOI: | 10.1016/j.cub.2014.02.060 |
| Popis: | Summary More than 2,000 C. elegans genes are targeted for RNA silencing by the mutator complex, a specialized small interfering RNA (siRNA) amplification module which is nucleated by the Q/N-rich protein MUT-16. The mutator complex localizes to Mutator foci adjacent to P granules at the nuclear periphery in germ cells [1]. Here, we show that the DEAD box RNA helicase smut-1 functions redundantly in the mutator pathway with its paralog mut-14 during RNAi. Mutations in both smut-1 and mut-14 also cause widespread loss of endogenous siRNAs. The targets of mut-14 and smut-1 largely overlap with the targets of other mutator class genes; however, the mut-14 smut-1 double mutant and the mut-16 mutant display the most dramatic depletion of siRNAs, suggesting that they act at a similarly early step in siRNA formation. mut-14 and smut-1 are predominantly expressed in the germline and, unlike other mutator class genes, are specifically required for RNAi targeting germline genes. A catalytically inactive, dominant-negative missense mutant of MUT-14 is RNAi defective in vivo; however, mutator complexes containing the mutant protein retain the ability to synthesize siRNAs in vitro. The results point to a role for mut-14 and smut-1 in initiating siRNA amplification in germ cell Mutator foci, possibly through the recruitment or retention of target mRNAs. |
| Databáze: | OpenAIRE |
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