Characterization of hnRNP K protein-RNA interactions
Characterization of hnRNP K protein-RNA interactions
复制标题
DOI：
10.1016/j.jmb.2004.07.099
复制
发表时间：
2004-09-24
影响因子：
5.6
通讯作者：
Ostrowski, J
中科院分区：
文献类型：
--
作者：
Klimek-Tomczak, K;Wyrwicz, LS;Ostrowski, J
异质核核糖核蛋白K蛋白是在几个亚细胞区室中发现的RNA结合蛋白，在这些亚细胞区室中，它被认为参与组成基因表达的多个过程的信号传导。K蛋白含有三个介导RNA结合的K同源（KH）结构域。我们使用了一系列的基因表达分析（SAGE）为基础的策略，酵母三杂交筛选，RNA下拉分析和计算分析，以表征K蛋白相关的RNA。我们证明，K蛋白相互作用与许多正义和反义核和线粒体转录通过直接和间接的结合。转录本与K蛋白的高度特异性直接结合由包含三个富含C的补丁的共有序列介导。结构分析表明，一个三管齐下的相互作用模型，其中每个三个KH域绑定的C-丰富的补丁之一。全基因组和酵母三杂交克隆分析表明，这些序列优先位于3'非翻译区，这是已知的调节mRNA的翻译和加工。(C)2004爱思唯尔有限公司保留所有权利。
The heterogeneous nuclear ribonucleoprotein K protein is an RNA-binding protein found in several subcellular compartments where it is thought to be involved in signaling multiple processes that compose gene expression. K protein contains three K homology (KH) domains that mediate RNA-binding. We used a serial analysis of gene expression (SAGE)-based strategy, yeast three-hybrid screen, RNA pull-down assays and computational analysis to characterize K protein-associated RNAs. We demonstrate that K protein interacts with many sense and antisense nuclear and mitochondrial transcripts through both direct and indirect binding. The highly specific direct binding of transcripts to K protein is mediated by a consensus sequence comprising three C-rich patches. Structural analysis suggests a three-prong interaction model whereby each of the three KH domains binds one of the C-rich patches. Genome-wide and yeast three-hybrid clone analysis revealed that these sequences are located preferentially in the 3' untranslated regions, which are known to regulate mRNA translation and processing. (C) 2004 Elsevier Ltd. All rights reserved.