Mapping genomic targets of DNA helicases by chromatin immunoprecipitation in Saccharomyces cerevisiae

Methods Mol Biol. 2010;587:113-26. doi: 10.1007/978-1-60327-355-8_8.

Abstract

DNA helicases utilize the energy of nucleotide hydrolysis to unwind the two annealed strands of the DNA helix and are involved in many aspects of DNA metabolism such as replication, recombination, and repair. Chromatin immunoprecipitation (ChIP) has been instrumental in determining the genomic targets of many DNA helicases and DNA helicase-containing complexes including the minichromosome maintenance (Mcm) proteins 2-7, the RecQ helicase Sgs1 as well as the Rvb1 and Rvb2 helicase-containing INO80 and SWR1 chromatin remodeling complexes. Here we describe a ChIP method that has been successfully used to map these proteins at chromosomal double-strand breaks and replication forks in the model organism Saccharomyces cerevisiae.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatin Immunoprecipitation / methods*
  • DNA / chemistry
  • DNA / metabolism*
  • DNA Helicases / chemistry
  • DNA Helicases / genetics
  • DNA Helicases / metabolism*
  • Polymerase Chain Reaction / methods
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • Saccharomyces cerevisiae Proteins
  • DNA
  • DNA Helicases