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Using Yeast as a Model Organism to Study the Functional Roles of Histone Acetylation in DNA Excision Repair

  • Amelia J. Hodges
  • Steven A. Roberts
  • John J. WyrickEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1983)

Abstract

Histone acetylation plays important roles in regulating DNA metabolic processes, including many DNA repair pathways. The nucleotide excision repair (NER) pathway is critical for removing bulky, helix-distorting DNA lesions, such as UV light-induced photoproducts, but the activity of this pathway is significantly inhibited when lesions reside in nucleosomes. Recent studies have indicated that histone acetyltransferase (HAT) activity may be induced in response to UV damage, in order to facilitate the repair of UV-induced lesions in chromatin. Budding yeast (Saccharomyces cerevisiae) is an important model system for studying the functional roles of histone acetylation and HATs in NER, due to the ease of genetically altering HAT activity or acetylated lysine residues in histones. Here, we describe protocols for measuring the repair of cyclobutane pyrimidine dimers (CPDs), the major UV-induced photoproduct, in yeast strains deficient in HAT activity, either due to gene deletion or rapid anchor-away depletion of the HAT enzyme. Methods for measuring CPD repair in bulk chromatin, as well as individual chromatin loci, are detailed below.

Keywords

Nucleotide excision repair Chromatin Histone Anchor-away Histone acetyltransferase Cyclobutane pyrimidine dimer UV light 

Notes

Acknowledgments

We thank Dr. Michael Smerdon, Dr. Peng Mao, and Dalton Plummer for helpful comments and suggestions. This research was supported by grants from NIEHS (R01ES002614 and R21ES027937).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Amelia J. Hodges
    • 1
  • Steven A. Roberts
    • 1
    • 2
  • John J. Wyrick
    • 1
    • 2
    Email author
  1. 1.School of Molecular BiosciencesWashington State UniversityPullmanUSA
  2. 2.Center of Reproductive BiologyWashington State UniversityPullmanUSA

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