小柯机器人

ChromID可鉴别染色质上的蛋白互作组
2020-03-03 16:45

瑞士苏黎世大学Tuncay Baubec研究组在研究中取得进展。他们建立了一种新工具ChromID,可在染色质标记处鉴别蛋白质的相互作用组。这一研究成果3月2日在线发表于《自然—生物技术》。

在这项研究中,研究人员使用天然蛋白质结构域作为模块化构建基石,来研发对H3K4、H3K9和H3K27位点DNA甲基化和组蛋白三甲基化选择性的工程染色质读取器(eCR)。

研究人员首先在小鼠胚胎干细胞中稳定表达eCRs并测量其亚核定位、基因组分布和组蛋白修饰结合偏好,证明了它们在活细胞中作为选择性染色质结合剂的作用。

通过将eCRs融合到生物素连接酶BASU中,研究人员建立了ChromID,这是一种基于邻近生物素识别染色质依赖性蛋白相互作用组的工具,并将其应用于小鼠干细胞中独特的染色质修饰。

使用合成的双修饰阅读器,研究人员还发现了由H3K4me3和H3K27me3标记的二价修饰启动子处蛋白质的组成。这些结果凸显了ChromID拥有获取染色质上蛋白质相互作用网络的能力。

据介绍,染色质修饰通过将蛋白质募集到基因组来调节基因组功能。但是,具有特异染色质修饰的蛋白质组成尚未得到充分表征。

附:英文原文

Title: ChromID identifies the protein interactome at chromatin marks

Author: Rodrigo Villaseor, Ramon Pfaendler, Christina Ambrosi, Stefan Butz, Sara Giuliani, Elana Bryan, Thomas W. Sheahan, Annika L. Gable, Nina Schmolka, Massimiliano Manzo, Jol Wirz, Christian Feller, Christian von Mering, Ruedi Aebersold, Philipp Voigt, Tuncay Baubec

Issue&Volume: 2020-03-02

Abstract: Chromatin modifications regulate genome function by recruiting proteins to the genome. However, the protein composition at distinct chromatin modifications has yet to be fully characterized. In this study, we used natural protein domains as modular building blocks to develop engineered chromatin readers (eCRs) selective for DNA methylation and histone tri-methylation at H3K4, H3K9 and H3K27 residues. We first demonstrated their utility as selective chromatin binders in living cells by stably expressing eCRs in mouse embryonic stem cells and measuring their subnuclear localization, genomic distribution and histone-modification-binding preference. By fusing eCRs to the biotin ligase BASU, we established ChromID, a method for identifying the chromatin-dependent protein interactome on the basis of proximity biotinylation, and applied it to distinct chromatin modifications in mouse stem cells. Using a synthetic dual-modification reader, we also uncovered the protein composition at bivalently modified promoters marked by H3K4me3 and H3K27me3. These results highlight the ability of ChromID to obtain a detailed view of protein interaction networks on chromatin.

DOI: 10.1038/s41587-020-0434-2

Source: https://www.nature.com/articles/s41587-020-0434-2

Nature Biotechnology:《自然—生物技术》,创刊于1996年。隶属于施普林格·自然出版集团,最新IF:68.164
官方网址:https://www.nature.com/nbt/
投稿链接:https://mts-nbt.nature.com/cgi-bin/main.plex


本期文章:《自然—生物技术》:Online/在线发表

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