ChAP-MS: A Method for Identification of Proteins and Histone Posttranslational Modifications at a Single Genomic Locus
The 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 fiel...
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doaj-82b2122c30b44c02b7bf302cc427b9392020-11-25T01:17:24ZengElsevierCell Reports2211-12472012-07-012119820510.1016/j.celrep.2012.06.019ChAP-MS: A Method for Identification of Proteins and Histone Posttranslational Modifications at a Single Genomic LocusStephanie D. Byrum0Ana Raman1Sean D. Taverna2Alan J. Tackett3University of Arkansas for Medical Sciences, Department of Biochemistry and Molecular Biology, Little Rock, AR 72205, USAJohns Hopkins School of Medicine, Department of Pharmacology and Molecular Sciences, Baltimore, MD 21205, USAJohns Hopkins School of Medicine, Department of Pharmacology and Molecular Sciences, Baltimore, MD 21205, USAUniversity of Arkansas for Medical Sciences, Department of Biochemistry and Molecular Biology, Little Rock, AR 72205, USA The 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. http://www.sciencedirect.com/science/article/pii/S2211124712001933 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Stephanie D. Byrum Ana Raman Sean D. Taverna Alan J. Tackett |
spellingShingle |
Stephanie D. Byrum Ana Raman Sean D. Taverna Alan J. Tackett ChAP-MS: A Method for Identification of Proteins and Histone Posttranslational Modifications at a Single Genomic Locus Cell Reports |
author_facet |
Stephanie D. Byrum Ana Raman Sean D. Taverna Alan J. Tackett |
author_sort |
Stephanie D. Byrum |
title |
ChAP-MS: A Method for Identification of Proteins and Histone Posttranslational Modifications at a Single Genomic Locus |
title_short |
ChAP-MS: A Method for Identification of Proteins and Histone Posttranslational Modifications at a Single Genomic Locus |
title_full |
ChAP-MS: A Method for Identification of Proteins and Histone Posttranslational Modifications at a Single Genomic Locus |
title_fullStr |
ChAP-MS: A Method for Identification of Proteins and Histone Posttranslational Modifications at a Single Genomic Locus |
title_full_unstemmed |
ChAP-MS: A Method for Identification of Proteins and Histone Posttranslational Modifications at a Single Genomic Locus |
title_sort |
chap-ms: a method for identification of proteins and histone posttranslational modifications at a single genomic locus |
publisher |
Elsevier |
series |
Cell Reports |
issn |
2211-1247 |
publishDate |
2012-07-01 |
description |
The 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.
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url |
http://www.sciencedirect.com/science/article/pii/S2211124712001933 |
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