RRC ID 11166
Author Doi N, Zenno S, Ueda R, Ohki-Hamazaki H, Ui-Tei K, Saigo K.
Title Short-interfering-RNA-mediated gene silencing in mammalian cells requires Dicer and eIF2C translation initiation factors.
Journal Curr Biol
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.
Volume 13(1)
Pages 41-6
Published 2003-1-8
DOI 10.1016/s0960-9822(02)01394-5
PII S0960-9822(02)01394-5
PMID 12526743
MeSH 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
IF 9.193
Times Cited 161