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A conserved siRNA-degrading RNase negatively regulates RNA interference in C. elegans
Scott Kennedy1, Duo Wang, Gary Ruvkun
1Department of Molecular Biology, Massachusetts General Hospital and Department of Genetics, Harvard Medical School, Boston, Massachusetts 02114, USA.
Nature
|February 13, 2004
Summary
The eri-1 gene mutation enhances RNA interference sensitivity by increasing short interfering RNA accumulation. This suggests ERI-1 acts as a negative regulator, controlling RNA interference duration and specificity.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- RNA interference (RNAi) is a conserved biological process where double-stranded RNA (dsRNA) triggers the degradation of homologous messenger RNA (mRNA).
- The efficiency of RNAi varies, with many mRNAs and nearly all Caenorhabditis elegans nervous system mRNAs being refractory to this process.
- The eri-1 gene was identified in a screen for C. elegans mutants exhibiting heightened sensitivity to dsRNA.
Purpose of the Study:
- To investigate the function of the eri-1 gene and its role in regulating RNA interference.
- To characterize the protein product of eri-1 and its enzymatic activity.
- To understand the implications of ERI-1 in controlling RNAi pathways.
Main Methods:
- Genetic screening in C. elegans to identify mutants with enhanced RNAi sensitivity.
- Molecular cloning and characterization of the eri-1 gene and its protein product.
- In vitro enzymatic assays using purified C. elegans ERI-1 and its human orthologue to assess siRNA degradation activity.
Main Results:
- Mutations in eri-1 lead to increased accumulation of short interfering RNAs (siRNAs) following dsRNA or siRNA exposure compared to wild-type animals.
- The eri-1 gene encodes an evolutionarily conserved protein with nucleic-acid-binding and exonuclease domains.
- Both C. elegans ERI-1 and its human orthologue demonstrate in vitro activity in degrading siRNAs.
- ERI-1 is primarily localized in the cytoplasm, with high expression in the gonad and specific neurons, suggesting tissue-specific regulation of RNAi.
Conclusions:
- ERI-1 functions as a negative regulator of RNA interference, likely by degrading siRNAs.
- The activity of ERI-1 may limit the duration, cell-type specificity, and endogenous functions of RNAi.
- Understanding ERI-1's role provides insights into the fine-tuning of RNAi pathways in different organisms and tissues.
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