ATP-dependent human RISC assembly pathways.

ATP-dependent human RISC assembly pathways.
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DOI:
10.1038/nsmb.1733
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发表时间:
2010-01
影响因子:
16.8
通讯作者:
--
中科院分区:
生物学1区
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rna诱导沉默复合体(RISC)的组装是小rna介导基因沉默的关键过程。在人类中,小干扰rna (sirna)和microRNAs (miRNAs)被整合到含有Argonaute (AGO)亚家族蛋白AGO 1 - 4的risc中。先前的研究已经提出,与果蝇的RISC组装途径不同,人类的RISC组装与切粒偶联,不依赖于ATP。在这里,我们通过仔细的重新检查表明,在人类中,RISC组装和切割是不耦合的,ATP极大地促进了小rna双链的RISC装载。此外,所有四种人类AGO蛋白对小rna双链表现出非常相似的结构偏好:中心错配促进RISC加载,种子或3 ' -mid(引导位置12-15)错配促进解绕。人类AGO蛋白的所有这些特征都与果蝇Ago2而非Ago2非常相似。
The assembly of RNA-induced silencing complex (RISC) is a key process in small RNA–mediated gene silencing. In humans, small interfering RNAs (siRNAs) and microRNAs (miRNAs) are incorporated into RISCs containing the Argonaute (AGO) subfamily proteins Ago1–4. Previous studies have proposed that, unlike Drosophila melanogaster RISC assembly pathways, human RISC assembly is coupled with dicing and is independent of ATP. Here we show by careful reexamination that, in humans, RISC assembly and dicing are uncoupled, and ATP greatly facilitates RISC loading of small-RNA duplexes. Moreover, all four human AGO proteins show remarkably similar structural preferences for small-RNA duplexes: central mismatches promote RISC loading, and seed or 3′-mid (guide position 12–15) mismatches facilitate unwinding. All these features of human AGO proteins are highly reminiscent of fly Ago1 but not fly Ago2.
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