Human TRBP and PACT directly interact with each other and associate with dicer to facilitate the production of small interfering RNA
Human TRBP and PACT directly interact with each other and associate with dicer to facilitate the production of small interfering RNA
复制标题
DOI：
10.1074/jbc.m611768200
复制
发表时间：
2007-06-15
影响因子：
4.8
通讯作者：
Jin, Dong-Yan
中科院分区：
文献类型：
--
作者：
Kok, Kin Hang;Ng, Ming-Him James;Jin, Dong-Yan
哺乳动物的Dicer与双链RNA结合蛋白TRBP或PACT相互作用，以介导RNA干扰和微小RNA加工。TRBP和PACT在结构上相关，但对PKR发挥相反的调节活性。目前尚不清楚Dicer是否同时需要TRBP和PACT。在此我们表明，TRBP在体外和哺乳动物细胞中都直接与PACT相互作用。TRBP和PACT与Dicer形成三元复合物，并促进Dicer产生小干扰RNA（siRNA）。在培养细胞中同时敲低TRBP和PACT会导致短发夹RNA介导的基因沉默受到显著抑制，但对siRNA介导的基因沉默没有影响，这表明TRBP和PACT主要在siRNA产生阶段发挥作用。综上所述，这些发现表明人类的TRBP和PACT直接相互作用，并与Dicer结合以刺激双链或短发夹RNA切割成siRNA。我们的工作极大地改变了目前关于在人类中产生siRNA和微小RNA的含Dicer复合物的组装和功能模型。
Mammalian Dicer interacts with double-stranded RNA-binding protein TRBP or PACT to mediate RNA interference and micro-RNA processing. TRBP and PACT are structurally related but exert opposite regulatory activities on PKR. It is not understood whether TRBP and PACT are simultaneously required for Dicer. Here we show that TRBP directly interacts with PACT in vitro and in mammalian cells. TRBP and PACT form a triple complex with Dicer and facilitate the production of small interfering RNA ( siRNA) by Dicer. Knockdown of both TRBP and PACT in cultured cells leads to significant inhibition of gene silencing mediated by short hairpin RNA but not by siRNA, suggesting that TRBP and PACT function primarily at the step of siRNA production. Taken together, these findings indicate that human TRBP and PACT directly interact with each other and associate with Dicer to stimulate the cleavage of double-stranded or short hairpin RNA to siRNA. Our work significantly alters the current model for the assembly and function of the Dicer-containing complex that generates siRNA and micro-RNA in human.