Genome-Wide Analysis of RNA-Protein Interactions in Plants
Genome-Wide Analysis of RNA-Protein Interactions in Plants
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DOI：
10.1007/978-1-60327-563-7_2
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发表时间：
2009-01-01
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影响因子：
--
通讯作者：
Barkan, Alice
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文献类型：
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作者：
Barkan, Alice
RNA-蛋白质相互作用通过其对基础基因表达机制和转录后过程的调节而深刻地影响生物体的发育和功能。预测的RNA结合蛋白（RBP）在植物中的库是特别大的，这表明在植物中的RNA-蛋白质相互作用组可能比后生动物中的RNA-蛋白质相互作用组更复杂和动态。为了剖析RNA-蛋白质相互作用网络，有必要确定每个RBP相互作用的RNA，并确定这些相互作用如何影响RNA发射和下游过程。RBP的天然RNA配体的鉴定仍然是一个挑战，但最近已经报道了几种用于分析与来自细胞提取物的特定RBP共纯化的RNA的高通量方法。本章回顾了在全基因组范围内定义RBPs的天然RNA配体的方法，并提供了一个协议的方法，已被用于这一目的的RBPs定位到叶绿体。
RNA-protein interactions profoundly impact organismal development and function through their contributions to the basal gene expression machineries and their regulation of post-transcriptional processes. The repertoire of predicted RNA binding proteins (RBPs) in plants is particularly large, suggesting that the RNA-protein interactome in plants may be more complex and dynamic even than that in metazoa. To dissect RNA-protein interaction networks, it is necessary to identify the RNAs with which each RBP interacts and to determine how those interactions influence RNA fire and downstream processes. Identification of the native RNA ligands of RBPs remains a challenge, but several high-throughput methods for the analysis of RNAs that copurify with specific RBPs from cell extract have been reported recently. This chapter reviews approaches for defining the native RNA ligands of RBPs on a genome-wide scale and provides a protocol for a method that has been used to this end for RBPs that localize to the chloroplast.