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PMID:12526743
| Citation |
Doi, N, Zenno, S, Ueda, R, Ohki-Hamazaki, H, Ui-Tei, K and Saigo, K (2003) Short-interfering-RNA-mediated gene silencing in mammalian cells requires Dicer and eIF2C translation initiation factors. Curr. Biol. 13:41-6 |
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| Abstract |
RNA interference (RNAi) is the process of long, double-stranded (ds), RNA-dependent posttranscriptional gene silencing (PTGS). In lower eukaryotes, dsRNA introduced into the cytoplasm is cleaved by the RNaseIII-like enzyme, Dicer, to 21-23 nt RNA (short interfering [si] RNA), which may serve as guide for target mRNA degradation. In mammals, long-dsRNA-dependent PTGS is applicable only to a limited number of cell types, whereas siRNA synthesized in vitro is capable of effectively inducing gene silencing in a wide variety of cells. Although biochemical and genetic analyses in lower eukaryotes showed that Dicer and some PIWI family member proteins are essential for long-dsRNA-dependent PTGS, little is known about the molecular mechanisms underlying siRNA-based PTGS. Here, we show that Dicer and eIF2C translation initiation factors belonging to the PIWI family (eIF2C1-4) play an essential role in mammalian siRNA-mediated PTGS, most probably through synergistic interactions. Immunoprecipitation experiments suggest that, in human and mouse cells, complex formation occurs between Dicer and eIF2C1 or 2 and that the PIWI domain of eIF2C is essential for the formation of this complex. |
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| Keywords |
3T3 Cells; Amino Acid Sequence; Animals; Argonaute Proteins; Endoribonucleases/genetics; Endoribonucleases/metabolism; Eukaryotic Initiation Factors; Gene Silencing/physiology; Genes, myc; HeLa Cells; Humans; Macromolecular Substances; Mammals/genetics; Mice; Molecular Sequence Data; Peptide Initiation Factors/genetics; Peptide Initiation Factors/metabolism; Precipitin Tests; Protein Biosynthesis/physiology; RNA Interference; RNA, Small Interfering/genetics; RNA, Small Interfering/physiology; Ribonuclease III |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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