用RNP圖譜分析RNA和蛋白質網絡確定RNA功能樞紐
作者：
小柯機器人發布時間：2020/10/22 14:38:16
美國北卡羅萊納大學Kevin M. Weeks研究小組的最新研究提出了RNP-MaP，用於分析活細胞中RNA-蛋白質相互作用網絡。 相關論文發表在2020年10月19日出版的《自然—生物技術》雜誌上。
研究人員設計了核糖核蛋白網絡分析突變剖析(RNP-MaP)，是一種活細胞化學探測策略，用於映射核苷酸解析度下多個蛋白質結合到單個核苷酸的合作互動。RNP-MaP使用異雙功能交聯劑，可將相互作用的蛋白質凍結在RNA上的原位置，然後通過通讀反轉錄和DNA測序將多個結合的蛋白質定位在單個RNA鏈上。
RNP-MaP揭示，核糖核酸酶P和RMRP，作為序列不同但結構相關聯的兩種非編碼RNA，共享RNP網絡且該網絡中心定義這些RNA的功能位點。RNP-MaP還確定了在老鼠和人類XIST長非編碼RNA之間保守的蛋白質相互作用網絡，並定義了其結合位點在XIST功能區域共定位並形成網絡的蛋白質群落。RNP-MaP實現了發現和有效驗證活細胞中長RNA和功能蛋白質相互作用網絡。
據悉，RNA蛋白質相互作用網絡管理許多生物過程，但很難全面檢測。
附：英文原文
Title: Analysis of RNA–protein networks with RNP-MaP defines functional hubs on RNA
Author: Chase A. Weidmann, Anthony M. Mustoe, Parth B. Jariwala, J. Mauro Calabrese, Kevin M. Weeks
Issue&Volume: 2020-10-19
Abstract: RNA–protein interaction networks govern many biological processes but are difficult to examine comprehensively. We devised ribonucleoprotein networks analyzed by mutational profiling (RNP-MaP), a live-cell chemical probing strategy that maps cooperative interactions among multiple proteins bound to single RNA molecules at nucleotide resolution. RNP-MaP uses a hetero-bifunctional crosslinker to freeze interacting proteins in place on RNA and then maps multiple bound proteins on single RNA strands by read-through reverse transcription and DNA sequencing. RNP-MaP revealed that RNase P and RMRP, two sequence-divergent but structurally related non-coding RNAs, share RNP networks and that network hubs define functional sites in these RNAs. RNP-MaP also identified protein interaction networks conserved between mouse and human XIST long non-coding RNAs and defined protein communities whose binding sites colocalize and form networks in functional regions of XIST. RNP-MaP enables discovery and efficient validation of functional protein interaction networks on long RNAs in living cells.
DOI: 10.1038/s41587-020-0709-7
Source: https://www.nature.com/articles/s41587-020-0709-7