Expanded RNA-binding activities of mammalian Argonaute 2.

Expanded RNA-binding activities of mammalian Argonaute 2.
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DOI:
10.1093/nar/gkp812
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发表时间:
2009-12
影响因子:
14.9
通讯作者:
Kiriakidou M
Kiriakidou M
中科院分区:
生物学2区
文献类型:
--
作者:
Tan GS;Garchow BG;Liu X;Yeung J;Morris JP 4th;Cuellar TL;McManus MT;Kiriakidou M

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哺乳动物的Argonaute 2(Ago2)蛋白与微小RNA(miRNAs)或小干扰RNA(siRNAs)结合,形成RNA诱导沉默复合物(RISCs/miRNPs)。在本研究中,我们对重组小鼠Ago2的RNA结合及核酸酶活性进行了表征。我们的研究表明,重组小鼠Ago2能高效地与miRNAs结合,形成有活性的RISC。令人惊讶的是,我们发现重组小鼠Ago2利用前体miRNAs或长的无结构单链RNA作为引导物形成有活性的RISC。此外,我们证明在体内,内源性人Ago2不依赖Dicer直接与前体miRNAs结合,并且在人细胞的细胞质和细胞核中都发现了Ago2:前体miRNA复合物。
Mammalian Argonaute 2 (Ago2) protein associates with microRNAs (miRNAs) or small interfering RNAs (siRNAs) forming RNA-induced silencing complexes (RISCs/miRNPs). In the present work, we characterize the RNA-binding and nucleolytic activity of recombinant mouse Ago2. Our studies show that recombinant mouse Ago2 binds efficiently to miRNAs forming active RISC. Surprisingly, we find that recombinant mouse Ago2 forms active RISC using pre-miRNAs or long unstructured single stranded RNAs as guides. Furthermore, we demonstrate that, in vivo, endogenous human Ago2 binds directly to pre-miRNAs independently of Dicer, and that Ago2:pre-miRNA complexes are found both in the cytoplasm and in the nucleus of human cells.
DOI: 10.1073/pnas.0808830105
发表时间: 2008-10-21
影响因子: 11.1
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