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Status |
Public on Jun 24, 2014 |
Title |
The Vasa homolog RDE-12 engages target mRNA and a secondary-Argonaute WAGO-1 to promote RNAi in C. elegans |
Organism |
Caenorhabditis elegans |
Experiment type |
Non-coding RNA profiling by high throughput sequencing
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Summary |
Argonaute proteins (AGOs) are key nuclease effectors of RNA interference (RNAi) [1]. Although purified AGOs can mediate a single round of target-RNA cleavage in vitro, accessory factors are required for siRNA loading and to achieve multiple-target turnover [2, 3]. To identify AGO co-factors we immunoprecipitated the C. elegans AGO WAGO-1, which engages amplified small RNAs during RNAi [4]. These studies identified a robust association between WAGO-1 and a conserved Vasa ATPase-related protein RDE-12. rde-12 mutants are deficient in RNAi including viral suppression, and fail to produce amplified secondary siRNAs and certain endogenous siRNAs (endo-siRNAs). RDE-12 co-localizes with WAGO-1 in germline P-granules and to peri-nuclear cytoplasmic foci in somatic cells. These findings and our genetic studies suggest that (i) RDE-12 is first recruited to target mRNAs by upstream AGOs (RDE-1 and ERGO-1) where it promotes small-RNA amplification and/or WAGO-1 loading, and that (ii) downstream of these events, RDE-12 forms an RNase-resistant (target mRNA-independent) complex with WAGO-1 that may scan for additional target mRNAs.
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Overall design |
Examine small RNA population changes in rde-12 mutants
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Contributor(s) |
Shirayama M, Stanney W, Gu W, Seth M, Mello C |
Citation(s) |
24684931 |
Submission date |
Jan 24, 2014 |
Last update date |
May 15, 2019 |
Contact name |
Craig Mello |
Organization name |
University of Massachusetts Medical School
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Department |
Program in Molecular Medicine
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Lab |
Craig Mello
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Street address |
368 Plantatoin Street, Suite AS5-2047
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City |
Worcester |
State/province |
MA |
ZIP/Postal code |
01605 |
Country |
USA |
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Platforms (1) |
GPL13776 |
Illumina Genome Analyzer IIx (Caenorhabditis elegans) |
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Samples (4)
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GSM1314315 |
total small RNAs in rde-12 with cpsf-2 RNAi food |
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Relations |
BioProject |
PRJNA236392 |
SRA |
SRP035672 |