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Chromatin affinity purification and quantitative mass spectrometry defining the interactome of histone modification patterns.

ABSTRACT: DNA and histone modifications direct the functional state of chromatin and thereby the readout of the genome. Candidate approaches and histone peptide affinity purification experiments have identified several proteins that bind to chromatin marks. However, the complement of factors that is recruited by individual and combinations of DNA and histone modifications has not yet been defined. Here, we present a strategy based on recombinant, uniformly modified chromatin templates used in affinity purification experiments in conjunction with SILAC-based quantitative mass spectrometry for this purpose. On the prototypic H3K4me3 and H3K9me3 histone modification marks we compare our method with a histone N-terminal peptide affinity purification approach. Our analysis shows that only some factors associate with both, chromatin and peptide matrices but that a surprisingly large number of proteins differ in their association with these templates. Global analysis of the proteins identified implies specific domains mediating recruitment to the chromatin marks. Our proof-of-principle studies show that chromatin templates with defined modification patterns can be used to decipher how the histone code is read and translated.

SUBMITTER: Nikolov M

PROVIDER: S-EPMC3226395 | biostudies-literature | 2011 Nov

REPOSITORIES: biostudies-literature

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Chromatin affinity purification and quantitative mass spectrometry defining the interactome of histone modification patterns.

Nikolov Miroslav M Stützer Alexandra A Mosch Kerstin K Krasauskas Andrius A Soeroes Szabolcs S Stark Holger H Urlaub Henning H Fischle Wolfgang W

Molecular & cellular proteomics : MCP 20110811 11

DNA and histone modifications direct the functional state of chromatin and thereby the readout of the genome. Candidate approaches and histone peptide affinity purification experiments have identified several proteins that bind to chromatin marks. However, the complement of factors that is recruited by individual and combinations of DNA and histone modifications has not yet been defined. Here, we present a strategy based on recombinant, uniformly modified chromatin templates used in affinity pur ...[more]

PMID: 21836164

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Dataset Information

Chromatin affinity purification and quantitative mass spectrometry defining the interactome of histone modification patterns.

Publications

Chromatin affinity purification and quantitative mass spectrometry defining the interactome of histone modification patterns.

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