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.
出处
《分子细胞生物学报》
CSCD
北大核心
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