Chromatin Immunoprecipitation to Detect DNA Replication and Repair Factors
| Autor: | Eishi Noguchi, Ken-ichi Noma, Mariana C. Gadaleta, Osamu Iwasaki, Chiaki Noguchi |
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| Rok vydání: | 2015 |
| Předmět: |
Cell Extracts
DNA Replication Genetics Chromatin Immunoprecipitation DNA Repair Cell Cycle DNA replication Eukaryotic DNA replication Biology Real-Time Polymerase Chain Reaction Pre-replication complex Antibodies Genomic Instability Article Cell biology Cross-Linking Reagents Replication factor C Control of chromosome duplication Minichromosome maintenance Schizosaccharomyces Origin recognition complex Schizosaccharomyces pombe Proteins Replication protein A |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493925957 |
| DOI: | 10.1007/978-1-4939-2596-4_12 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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