In vivo binding of recombination proteins to non-DSB DNA lesions and to replication forks
AffiliationDepartment of Cell and Chemical Biology, Leiden University Medical Center, Leiden, The Netherlands.
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AbstractHomologous recombination (HR) has been extensively studied in response to DNA double-strand breaks (DSBs). In contrast, much less is known about how HR deals with DNA lesions other than DSBs (e.g., at single-stranded DNA) and replication forks, despite the fact that these DNA structures are associated with most spontaneous recombination events. A major handicap for studying the role of HR at non-DSB DNA lesions and replication forks is the difficulty of discriminating whether a recombination protein is associated with the non-DSB lesion per se or rather with a DSB generated during their processing. Here, we describe a method to follow the in vivo binding of recombination proteins to non-DSB DNA lesions and replication forks. This approach is based on the cleavage and subsequent electrophoretic analysis of the target DNA by the recombination protein fused to the micrococcal nuclease.
CitationGonzalez-Prieto R, Cabello-Lobato MJ, Prado F. In Vivo Binding of Recombination Proteins to Non-DSB DNA Lesions and to Replication Forks. Methods Mol Biol. 2021;2153:447-58.
JournalMethods in Molecular Biology
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