Journal
NUCLEIC ACIDS RESEARCH
Volume 41, Issue 20, Pages -Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkt822
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Funding
- National Institutes of Health (NIH) [R01GM106024, P30GM103450, P20GM103429, UL1TR000039]
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Here, we describe an approach to isolate native chromatin sections without genomic engineering for label-free proteomic identification of associated proteins and histone post-translational modifications. A transcription activator-like (TAL) protein A fusion protein was designed to recognize a unique site in the yeast GAL1 promoter. The TAL-PrA fusion enabled chromatin affinity purification (ChAP) of a small section of native chromatin upstream from the GAL1 locus, permitting mass spectrometric (MS) identification of proteins and histone post-translational modifications regulating galactose-induced transcription. This TAL-ChAP-MS approach allows the biochemical isolation of a specific native genomic locus for proteomic studies and will provide for unprecedented objective insight into protein and epigenetic mechanisms regulating site-specific chromosome metabolism.
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