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Chromatin Immunoprecipitation to Detect DNA Replication and Repair Factors
Book chapter   Open access

Chromatin Immunoprecipitation to Detect DNA Replication and Repair Factors

Mariana C Gadaleta, Osamu Iwasaki, Chiaki Noguchi, Ken-Ichi Noma and Eishi Noguchi
DNA Replication
2015
DOI: https://doi.org/10.1007/978-1-4939-2596-4_12
PMID: 25916713
url
https://doi.org/10.1007/978-1-4939-2596-4_12View
Published, Version of Record (VoR) Open

Abstract

Chromatin immunoprecipitation DNA replication Difficult to replicate Replication fork Recombination Rad52 DNA damage Protein–DNA cross-linking ChIP DNA repair Quantitative PCR
DNA replication is tightly coupled with DNA repair processes in order to preserve genomic integrity. During DNA replication, the replication fork encounters a variety of obstacles including DNA damage/adducts, secondary structures, and programmed fork-blocking sites, which are all difficult to replicate. The replication fork also collides with the transcription machinery, which shares the template DNA with the replisome complex. Under these conditions, replication forks stall, causing replication stress and/or fork collapse, ultimately leading to genomic instability. The mechanisms to overcome these replication problems remain elusive. Therefore, it is important to investigate how DNA repair and replication factors are recruited and coordinated at chromosomal regions that are difficult to replicate. In this chapter, we describe a chromatin immunoprecipitation method to locate proteins required for DNA repair during DNA replication in the fission yeast Schizosaccharomyces pombe. This method can also easily be adapted to study replisome components or chromatin-associated factors.

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