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Single-molecule decoding of combinatorially modified nucleosomes.


ABSTRACT: Different combinations of histone modifications have been proposed to signal distinct gene regulatory functions, but this area is poorly addressed by existing technologies. We applied high-throughput single-molecule imaging to decode combinatorial modifications on millions of individual nucleosomes from pluripotent stem cells and lineage-committed cells. We identified definitively bivalent nucleosomes with concomitant repressive and activating marks, as well as other combinatorial modification states whose prevalence varies with developmental potency. We showed that genetic and chemical perturbations of chromatin enzymes preferentially affect nucleosomes harboring specific modification states. Last, we combined 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.

SUBMITTER: Shema E 

PROVIDER: S-EPMC4904710 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature

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Single-molecule decoding of combinatorially modified nucleosomes.

Shema Efrat E   Jones Daniel D   Shoresh Noam N   Donohue Laura L   Ram Oren O   Bernstein Bradley E BE  

Science (New York, N.Y.) 20160501 6286


Different combinations of histone modifications have been proposed to signal distinct gene regulatory functions, but this area is poorly addressed by existing technologies. We applied high-throughput single-molecule imaging to decode combinatorial modifications on millions of individual nucleosomes from pluripotent stem cells and lineage-committed cells. We identified definitively bivalent nucleosomes with concomitant repressive and activating marks, as well as other combinatorial modification s  ...[more]

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