Argonaute Utilization for miRNA Silencing Is Determined by Phosphorylation-Dependent Recruitment of LIM-Domain-Containing Proteins.
Argonaute Utilization for miRNA Silencing Is Determined by Phosphorylation-Dependent Recruitment of LIM-Domain-Containing Proteins.
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DOI：
10.1016/j.celrep.2017.06.027
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发表时间：
2017-07-05
期刊：
影响因子：
8.8
通讯作者：
Sharp TV
中科院分区：
文献类型：
--
作者：
Bridge KS;Shah KM;Li Y;Foxler DE;Wong SCK;Miller DC;Davidson KM;Foster JG;Rose R;Hodgkinson MR;Ribeiro PS;Aboobaker AA;Yashiro K;Wang X;Graves PR;Plevin MJ;Lagos D;Sharp TV
作为microrna诱导沉默复合体（miRISC）的核心成分，Argonaute （AGO）蛋白与TNRC6蛋白相互作用，募集翻译抑制/mRNA不稳定的其他效应物。在这里，我们发现LIMD1通过在不同的界面同时结合两种蛋白来协调含有miRISC复合物的AGO-TNRC6的组装。Akt3磷酸化AGO2 Ser 387诱导LIMD1结合，进而使AGO2与TNRC6A和下游效应物DDX6相互作用。该丝氨酸在AGO1和4中的保守性表明该机制可能是AGO功能和miRISC组装的基本要求。crispr - cas9介导的LIMD1敲除后，AGO2 miRNA沉默功能丧失，miRNA沉默依赖于AGO2与LIMD1家族成员WTIP形成的复合物。由于在相互作用界面上存在谷氨酸残基（E390），因此AGO3可以与LIMD1、AJUBA和WTIP结合，而不受Akt信号的影响。AGO1、2和4以磷酸化依赖的方式招募LIMD1，在HeLa细胞中，当LIMD1消失时，AGO2向AGO3转换发生。AGO3- wtip以不依赖akt3的方式取代AGO2-LIMD1进行miRNA沉默，Argonaute （AGO）蛋白通过形成microrna诱导的沉默复合体（miRISC）介导转录后基因沉默。Bridge等人发现含有lim结构域的蛋白通过磷酸化依赖机制对miRISC的形成至关重要。这对转录后基因沉默至关重要，并揭示了miRISC功能是通过“AGO开关”维持的。
As core components of the microRNA-induced silencing complex (miRISC), Argonaute (AGO) proteins interact with TNRC6 proteins, recruiting other effectors of translational repression/mRNA destabilization. Here, we show that LIMD1 coordinates the assembly of an AGO-TNRC6 containing miRISC complex by binding both proteins simultaneously at distinct interfaces. Phosphorylation of AGO2 at Ser 387 by Akt3 induces LIMD1 binding, which in turn enables AGO2 to interact with TNRC6A and downstream effector DDX6. Conservation of this serine in AGO1 and 4 indicates this mechanism may be a fundamental requirement for AGO function and miRISC assembly. Upon CRISPR-Cas9-mediated knockout of LIMD1, AGO2 miRNA-silencing function is lost and miRNA silencing becomes dependent on a complex formed by AGO3 and the LIMD1 family member WTIP. The switch to AGO3 utilization occurs due to the presence of a glutamic acid residue (E390) on the interaction interface, which allows AGO3 to bind to LIMD1, AJUBA, and WTIP irrespective of Akt signaling. LIMD1 is necessary for the Akt3-dependent assembly of an AGO-TNRC6A functional miRISC AGO1, 2, and 4 recruit LIMD1 in a phospho-dependent manner AGO2 to AGO3 switching occurs upon LIMD1 ablation in HeLa cells AGO3-WTIP replaces AGO2-LIMD1 for miRNA silencing in an Akt3-independent manner Argonaute (AGO) proteins mediate post-transcriptional gene silencing through formation of the microRNA-induced silencing complex (miRISC). Bridge et al. identify LIM-domain-containing proteins as essential for miRISC formation through a phosphorylation-dependent mechanism. This is critical for post-transcriptional gene silencing and reveals that miRISC functionality is maintained by “AGO switching.”
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