ChAP-MS: A method for identification of proteins and histone posttranslational modifications at a single genomic locus

Stephanie D. Byrum, Ana Raman, Sean Dixon Taverna, Alan J. Tackett

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)198-205
Number of pages8
JournalCell Reports
Volume2
Issue number1
DOIs
StatePublished - Jul 26 2012

Fingerprint

Histone Code
Post Translational Protein Processing
Histones
Chromatin
Purification
Mass spectrometry
Mass Spectrometry
Epigenomics
Proteins
Chromatin Immunoprecipitation
Proteome
Base Pairing
Proteomics
Transcription

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

ChAP-MS : A method for identification of proteins and histone posttranslational modifications at a single genomic locus. / Byrum, Stephanie D.; Raman, Ana; Taverna, Sean Dixon; Tackett, Alan J.

In: Cell Reports, Vol. 2, No. 1, 26.07.2012, p. 198-205.

Research output: Contribution to journalArticle

@article{aeafb63d646248858fe5574f920c7c29,
title = "ChAP-MS: A method for identification of proteins and histone posttranslational modifications at a single genomic locus",
abstract = "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.",
author = "Byrum, {Stephanie D.} and Ana Raman and Taverna, {Sean Dixon} and Tackett, {Alan J.}",
year = "2012",
month = "7",
day = "26",
doi = "10.1016/j.celrep.2012.06.019",
language = "English (US)",
volume = "2",
pages = "198--205",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "1",

}

TY - JOUR

T1 - ChAP-MS

T2 - A method for identification of proteins and histone posttranslational modifications at a single genomic locus

AU - Byrum, Stephanie D.

AU - Raman, Ana

AU - Taverna, Sean Dixon

AU - Tackett, Alan J.

PY - 2012/7/26

Y1 - 2012/7/26

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84864308665&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84864308665&partnerID=8YFLogxK

U2 - 10.1016/j.celrep.2012.06.019

DO - 10.1016/j.celrep.2012.06.019

M3 - Article

C2 - 22840409

AN - SCOPUS:84864308665

VL - 2

SP - 198

EP - 205

JO - Cell Reports

JF - Cell Reports

SN - 2211-1247

IS - 1

ER -