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Given the frequent misregulation of chromatin in cancer, it is important to understand the cellular mechanisms that regulate chromatin structure. However, systematic screening for epigenetic regulators is challenging and often relies on laborious assays or indirect reporter read-outs. Here we describe a strategy, Epi-ID, to directly assess chromatin status in thousands of mutants. In Epi-ID, chromatin status on DNA barcodes is interrogated by chromatin immunoprecipitation followed by deep sequencing, allowing for quantitative comparison of many mutants in parallel. Screening of a barcoded yeast knock-out collection for regulators of histone H3K79 methylation by Dot1 identified all known regulators as well as novel players and processes. These include histone deposition, homologous recombination, and adenosine kinase, which influences the methionine cycle. Gcn5, the acetyltransferase within the SAGA complex, was found to regulate histone methylation and H2B ubiquitination. The concept of Epi-ID is widely applicable and can be readily applied to other chromatin features.
Pubmed ID: 27922451 RIS Download
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View all literature mentionsThis polyclonal targets HIST1H3A
View all literature mentionsThis monoclonal targets Yeast 3-phosphoglycerate kinase (PGK)
View all literature mentionsThis polyclonal targets Sir2 (yN-19)
View all literature mentionsThis polyclonal targets H3K36me3
View all literature mentionsThis monoclonal targets dimethyl-Histone H3 (Lys79) clone NL59
View all literature mentionsThis polyclonal targets Histone H3 (tri methyl K4)
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View all literature mentionsSystem that classifies genes by their functions, using published scientific experimental evidence and evolutionary relationships to predict function even in absence of direct experimental evidence. Orthologs view is curated orthology relationships between genes for human, mouse, rat, fish, worm, and fly.
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