Article ID Journal Published Year Pages File Type
2041044 Cell Reports 2012 8 Pages PDF
Abstract

SummaryThe field of epigenomics has been transformed by chromatin immunoprecipitation approaches that provide for the localization of a defined protein or posttranslationally modified protein to specific chromosomal sites. While these approaches have helped us conceptualize epigenetic mechanisms, the field has been limited by the inability to define features such as the proteome and histone modifications at a specific genomic locus in an unbiased manner. We developed an unbiased approach whereby a unique native genomic locus was isolated, which was followed by high-resolution proteomic identification of specifically associated proteins and histone posttranslational modifications. This chromatin affinity purification with mass spectrometry (ChAP-MS) technique was used to specifically enrich a ∼1,000 base pair section of GAL1 chromatin under transcriptionally active and repressive conditions, as well as to identify the specifically bound proteins and histone posttranslational modifications. ChAP-MS should yield insight into the regulatory mechanisms of transcription and help identify factors that epigenetically control chromatin function.

Graphical AbstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► ChAP-MS allows the isolation of a ∼1 kb section of native chromatin ► Proteins and histone posttranslational modifications are identified by mass spectrometry ► ChAP-MS allows the isolation of a single locus for proteomic analysis ► ChAP-MS will provide insight into epigenetic regulatory mechanisms

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Life Sciences Agricultural and Biological Sciences Agricultural and Biological Sciences (General)
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