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首页> 外文期刊>Current Biology: CB >Short-interfering-RNA-mediated gene silencing in mammalian cells requires dicer and eIF2C translation initiation factors
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Short-interfering-RNA-mediated gene silencing in mammalian cells requires dicer and eIF2C translation initiation factors

机译：哺乳动物细胞中短干扰RNA介导的基因沉默需要切酶和eIF2C翻译起始因子

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摘要

RNA interference (RNAi) is the process of long, double-stranded (ds), RNA-dependent posttranscriptional gene silencing (PTGS) [1]. In lower eukaryotes, dsRNA introduced into the cytoplasm is cleaved by the RNaselll-like enzyme, Dicer, to 21-23 nt RNA (short interfering [si] RNA), which may serve as guide for target mRNA degradation [2]. In mammals, long-dsRNA-dependent PTGS is applicable only to a limited number of cell types [3-7], whereas siRNA synthesized in vitro is capable of effectively inducing gene silencing in a wide variety of cells [8]. Although biochemical and genetic analyses in lower eukaryotes; showed that Dicer and some PIWI family member proteins are essential for long-dsRNA-dependent PTGS [9-11], 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.
机译：RNA干扰（RNAi）是长双链（ds），依赖RNA的转录后基因沉默（PTGS）的过程[1]。在低等真核生物中，引入到细胞质中的dsRNA被RNaselll样酶Dicer切割成21-23 nt RNA（短干扰[si] RNA），可作为靶标mRNA降解的指导[2]。在哺乳动物中，长dsRNA依赖性的PTGS仅适用于有限数量的细胞类型[3-7]，而体外合成的siRNA能够在多种细胞中有效诱导基因沉默[8]。尽管在低等真核生物中进行了生化和遗传分析;结果表明，Dicer和一些PIWI家族成员蛋白对于长dsRNA依赖的PTGS是必不可少的[9-11]，对基于siRNA的PTGS的分子机制了解甚少。在这里，我们显示属于PIWI家族的Dicer和eIF2C翻译起始因子（eIF2C1-4）在哺乳动物siRNA介导的PTGS中起着至关重要的作用，很可能是通过协同相互作用。免疫沉淀实验表明，在人和小鼠细胞中，在Dicer和eIF2C1或2之间发生复合物形成，而eIF2C的PIWI结构域对于这种复合物的形成至关重要。

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