Single-molecule decoding of combinatorially modified nucleosomes.
Single-molecule decoding of combinatorially modified nucleosomes.
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DOI：
10.1126/science.aad7701
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发表时间：
2016-05-06
期刊：
影响因子：
--
通讯作者：
Bernstein BE
中科院分区：
文献类型：
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作者：
Shema E;Jones D;Shoresh N;Donohue L;Ram O;Bernstein BE
已提出组蛋白修饰的不同组合来表示不同的基因调控功能，但现有技术很难解决。在这里，我们应用高通量单分子成像来解码来自多能干细胞和谱系定向细胞的数百万个单个核小体的组合修饰。我们明确地鉴定出具有抑制和激活标记的二价核小体，以及其他组合修饰状态，其普遍程度随发育效力而变化。我们发现染色质酶的遗传和化学扰动都会优先影响具有特定修饰状态的核小体。最后，我们将该蛋白质组学平台与单分子DNA测序技术相结合，同时确定单个核小体的修饰状态和基因组位置。这种单分子技术有潜力解决染色质生物学和表观遗传调控的基本问题。
Different combinations of histone modifications have been proposed to signal distinct gene regulatory functions, but are poorly addressed by existing technologies. Here, we apply high-throughput single-molecule imaging to decode combinatorial modifications on millions of individual nucleosomes from pluripotent stem cells and lineage-committed cells. We identify definitively bivalent nucleosomes with concomitant repressive and activating marks, as well as other combinatorial modification states whose prevalence varies with developmental potency. We show that genetic and chemical perturbations of chromatin enzymes both preferentially affect nucleosomes harboring specific modification states. Finally, we combine this proteomic platform with single-molecule DNA sequencing technology to simultaneously determine the modification states and genomic positions of individual nucleosomes. This single-molecule technology has the potential to address fundamental questions in chromatin biology and epigenetic regulation.