Dicer蛋白与Wig-1蛋白的相互作用的研究  

Dicer INTERACTS WITH Wig-1 PROTEIN

作　　者：黄建[1,2] 王海波[1] 姚玲[1] 郑新民[1] Olof Radmark 

机构地区：[1]上海交通大学医学院生物化学与分子生物学教研室 [2]Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, S-17177, Sweden

出　　处：《分子细胞生物学报》2008年第5期376-382,共7页Journal of Molecular Cell Biology

基　　金：The Swedish Medical Research Council(03X-217);The European Union(QLG1-CT-2001-01521)资助

摘　　要：Dicer是RNA干扰中起核心作用的一种dsRNase,Wig-1是受p53诱导的、与dsRNA特异结合的一种锌指蛋白。本研究初步证实了Dicer蛋白与Wig-1蛋白存在相互作用。GST pull-down实验表明Dicer蛋白能与GST-Wig-1结合,而Wig-1蛋白能竞争性干扰Dicer与GST-eIF2C1的结合;Far-western分析证实Dicer蛋白可以与结合在硝酸纤维素膜上的GST-Wig-1蛋白结合;进一步发现Wig-1能有效促进重组Dicer的dsRNase酶切活性。这些结果揭示Dicer蛋白和Wig-1蛋白很可能通过相互作用共同参与了dsRNA调控相关的生理过程。Dicer is a key dsRNase in RNA interference, and Wig-1 is a p53-induced zinc finger protein with dsRNA-binding activity. In this report, we demonstrated a direct interaction between Dicer and Wig-1. Dicer associated with Wig-1, which was expressed as a glutathione Stransferase fusion protein, in a dose-dependent manner by GST pull-down assay. In a reciprocal experiment, Wig-1 appeared to compete with GST-eIF2C1 for the binding of Dicer. Far-western analysis showed that Dicer could bind to GST-Wig-1 directly. Furthermore, it was found that Wig-1 could increase the dsRNase activity of Dicer in vitro. Our findings raise the possibility that Dicer and Wig-1 may play a common role in dsRNA-related gene regulation.

关 键 词：DICER Wig-1 GST pull—down实验 Far—western分析 dsRNase酶切活性 

分 类 号：Q51[生物学—生物化学]