The Analysis of Recombination-Dependent Processing of Blocked Replication Forks by Bidimensional Gel Electrophoresis.

Autor: Kramarz K; Institut Curie, CNRS, UMR3348, PSL Research University, Orsay, France.; Institut Curie, Paris-Saclay University, Unite Mixte de Recherche, Centre National de la Recherche Scientifique, Orsay, France.; Genome Integrity, RNA and Cancer Unit (UMR3348), Equipe Labelisée Ligue, Institut Curie - Research Center, Orsay, France., Saada AA; School of Biological Sciences, Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA, USA., Lambert SAE; Institut Curie, CNRS, UMR3348, PSL Research University, Orsay, France. sarah.lambert@curie.fr.; Institut Curie, Paris-Saclay University, Unite Mixte de Recherche, Centre National de la Recherche Scientifique, Orsay, France. sarah.lambert@curie.fr.; Genome Integrity, RNA and Cancer Unit (UMR3348), Equipe Labelisée Ligue, Institut Curie - Research Center, Orsay, France. sarah.lambert@curie.fr.
Jazyk: angličtina
Zdroj: Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2153, pp. 365-381.
DOI: 10.1007/978-1-0716-0644-5_25
Abstrakt: The perturbation of the DNA replication process is a threat to genome stability and is an underlying cause of cancer development and numerous human diseases. It has become central to understanding how stressed replication forks are processed to avoid their conversion into fragile and pathological DNA structures. The engineering of replication fork barriers (RFBs) to conditionally induce the arrest of a single replisome at a defined locus has made a tremendous impact in our understanding of replication fork processing. Applying the bidimensional gel electrophoresis (2DGE) technique to those site-specific RFBs allows the visualization of replication intermediates formed in response to replication fork arrest to investigate the mechanisms ensuring replication fork integrity. Here, we describe the 2DGE technique applied to the site-specific RTS1-RFB in Schizosaccharomyces pombe and explain how this approach allows the detection of arrested forks undergoing nascent strands resection.
Databáze: MEDLINE
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