Functional Dissection of the Human TNRC6 (GW182-Related) Family of Proteins
Functional Dissection of the Human TNRC6 (GW182-Related) Family of Proteins
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DOI：
10.1128/mcb.00380-09
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发表时间：
2009-08-01
影响因子：
5.3
通讯作者：
Shiekhattar, Ramin
中科院分区：
文献类型：
--
作者：
Baillat, David;Shiekhattar, Ramin
Argonaute（Ago）蛋白通过其与小RNA的缔合在RNA干扰的效应子步骤中执行关键功能。TNRC 6（含三核苷酸重复序列6）蛋白家族已显示与哺乳动物细胞中的Agos稳定缔合。在这里，我们描述了TNRC 6 B-和TNRC 6C-含有复合物的分离和功能表征。我们表明，TNRC 6 B和TNRC 6C蛋白与所有四种已经加载了microRNA的人Agos相关。TNRC 6 B蛋白的详细结构域分析表明，Ago结合和P体定位需要该蛋白的不同结构域。使用响应于通过MS 2结合结构域拴系的TNRC 6 B的报告构建体进行的功能分析表明，翻译沉默既不需要Ago结合结构域也不需要P体定位结构域。相反，含有RNA识别基序的C-末端结构域在由TNRC 6 B蛋白介导的沉默中起关键作用。
Argonaute (Ago) proteins through their association with small RNAs perform a critical function in the effector step of RNA interference. The TNRC6 (trinucleotide repeat containing 6) family of proteins have been shown to stably associate with Agos in mammalian cells. Here, we describe the isolation and functional characterization of TNRC6B- and TNRC6C-containing complexes. We show that TNRC6B and TNRC6C proteins associate with all four human Agos which are already loaded with microRNAs. Detailed domain analysis of TNRC6B protein indicated that distinct domains of the protein are required for Ago binding and P-body localization. Functional analysis using reporter constructs responsive to TNRC6B tethered through an MS2-binding domain indicates that neither the Ago-binding nor the P-body localization domains are required for translational silencing. In contrast, the C-terminal domain containing the RNA recognition motif plays a critical role in the silencing mediated by the TNRC6B protein.