PDF(Pubmed)
Abstract：
Interactions between double-stranded RNA (dsRNA) and proteins play an important role in cellular homeostasis by regulating the editing, stability, and splicing of intracellular RNA. The identification of dsRNA-binding proteins (dsRBPs) is key; however, it has long been challenging to purify dsRBPs from cells. In this study, we developed a novel method, dsRBPC (dsRNA-binding protein capture), to purify cellular dsRBPs based on classic phase separation purification procedures. A global dsRNA-binding proteome of LLC-PK1 cells was obtained, and we identified 1326 dsRBPs, including 1303 putative novel dsRBPs. Functional analyses suggested that these enriched dsRBPs are mainly associated with rRNA processing, RNA splicing, transcriptional regulation, and nucleocytoplasmic transport. We also found that the ARM (armadillo/beta-catenin-like repeats) motif is a previously unknown dsRNA-binding domain, as demonstrated by biochemical experiments. Collectively, this study provides a useful approach for dsRBP identification and the discovery of a global dsRNA-binding proteome to comprehensively map the dsRNA - protein interaction network.
摘要：
双链RNA(dsRNA)和蛋白质之间的相互作用通过调节编辑在细胞内稳态中发挥重要作用,稳定性,和细胞内RNA的剪接。dsRNA结合蛋白(dsRBP)的鉴定是关键;然而,从细胞中纯化dsRBP一直是具有挑战性的。在这项研究中,我们开发了一种新的方法，dsRBPC（dsRNA结合蛋白捕获），基于经典的相分离纯化程序纯化细胞dsRBP。获得了LLC-PK1细胞的全局dsRNA结合蛋白质组，我们确定了1326个dsRBP，包括1303个推定的新型dsRBP。功能分析表明，这些富集的dsRBP主要与rRNA加工相关，RNA剪接,转录调控，和核质运输。我们还发现ARM（Armadillo/β-catenin-like重复序列）基序是以前未知的dsRNA结合域，正如生化实验所证明的。总的来说，本研究为dsRBP鉴定和发现全局dsRNA结合蛋白质组提供了一种有用的方法，以全面绘制dsRNA-蛋白质相互作用网络。
