In Vitro Analysis of Ribonucleoprotein Complex Remodeling and Disassembly.
In Vitro Analysis of Ribonucleoprotein Complex Remodeling and Disassembly.
复制标题
核糖核蛋白复合物重塑和拆卸的体外分析。
DOI：
10.1007/978-1-4939-3591-8_7
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Lou,Hua
中科院分区：
文献类型：
--
作者：
Zhou,Hua-Lin;Lou,Hua
核糖核蛋白复合物在基因表达中起着重要作用。它们的组装和分解控制着mRNA分子的命运。在这里，我们描述了一种方法，检查RNP的重塑和拆卸。该方法的一个独特方面是在具有或不具有所需修饰的HeLa细胞中产生感兴趣的RNA结合蛋白（RBP），并且在细胞提取物中用合成的RNA寡核苷酸组装RNP。我们使用这种方法来研究RBP的泛素化如何影响其结合其RNA靶标的能力。
Ribonucleoprotein (RNP) complexes play essential roles in gene expression. Their assembly and disassembly control the fate of mRNA molecules. Here, we describe a method that examines the remodeling and disassembly of RNPs. One unique aspect of this method is that the RNA-binding proteins (RBPs) of interest are produced in HeLa cells with or without the desired modification and the RNP is assembled in cellular extracts with synthetic RNA oligonucleotides. We use this method to investigate how ubiquitination of an RBP affects its ability to bind its RNA target.