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Chromatin Immunoprecipitation to Investigate Origin Association of Replication Factors in Mammalian Cells
Book chapter   Open access

Chromatin Immunoprecipitation to Investigate Origin Association of Replication Factors in Mammalian Cells

Adam R Leman and Eishi Noguchi
Cell Cycle Control, pp 539-547
14 May 2014
PMID: 24906335
url
https://doi.org/10.1007/978-1-4939-0888-2_30View
Published, Version of Record (VoR) Open

Abstract

Origin Chromatin immunoprecipitation Replication fork Timeless Fork protection complex c-Myc origin Replisome ChIP Cross-link
A variety of DNA-binding proteins regulate DNA transactions including DNA replication and DNA damage response. To initiate DNA replication in S phase of the cell cycle, numerous replication proteins must be recruited to the replication origin in order to unwind and synthesize DNA. Some replication factors stay at the origin, while replisome components move with the replication fork. When the replisome encounters DNA damage or other issues during DNA replication, the replication fork stalls and accumulates single-stranded DNA that triggers the ATR-dependent replication checkpoint, in order to slow down S phase and arrest the cell cycle at the G2–M transition. It is also possible that replication forks collapse, leading to double-strand breaks that recruit various DNA damage response proteins to activate cell cycle checkpoints and DNA repair pathways. Therefore, defining the localization of DNA transaction factors during the cell cycle should provide important insights into mechanistic understanding of DNA replication and its related processes. In this chapter, we describe a chromatin immunoprecipitation method to locate replisome components at replication origins in human cells.

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Collaboration types
Domestic collaboration
Web of Science research areas
Biochemical Research Methods
Biochemistry & Molecular Biology
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