Measuring Dynamic Changes in Histone Modifications and Nucleosome Density during Activated Transcription in Budding Yeast

  • Chhabi K. Govind
  • Daniel Ginsburg
  • Alan G. Hinnebusch
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 833)

Abstract

Chromatin immunoprecipitation is widely utilized to determine the in vivo binding of factors that regulate transcription. This procedure entails formaldehyde-mediated cross-linking of proteins and isolation of soluble chromatin followed by shearing. The fragmented chromatin is subjected to immunoprecipitation using antibodies against the protein of interest and the associated DNA is identified using quantitative PCR. Since histones are posttranslationally modified during transcription, this technique can be effectively used to determine the changes in histone modifications that occur during transcription. In this paper, we describe a detailed methodology to determine changes in histone modifications in budding yeast that takes into account reductions in nucleosome.

Key words

Activated transcription RNA polymerase II Histone modifications Acetylation Gcn4 Gal4 Saccharomyces cerevisiae Histone acetyltransferase Histone deacetylase complexes 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Chhabi K. Govind
    • 1
  • Daniel Ginsburg
    • 2
  • Alan G. Hinnebusch
    • 3
  1. 1.Department of Biological SciencesOakland UniversityRochesterUSA
  2. 2.Department of Biomedical SciencesLong Island UniversityBrookvilleUSA
  3. 3.Laboratory of Gene Regulation and DevelopmentEunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of HealthBethesdaUSA

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