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

Methods in Molecular Biology
Jennifer Cobb, Haico van Attikum

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.

Related Concepts

DNA, Double-Stranded
ATP-Dependent DNA Helicases
Saccharomyces cerevisiae
Nested Polymerase Chain Reaction
Saccharomyces cerevisiae Proteins
Chromatin Immunoprecipitation
DNA
Genome
Nucleotides
Recombination, Genetic

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