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Título

In Vivo Binding of Recombination Proteins to Non-DSB DNA Lesions and to Replication Forks

AutorGonzález-Prieto, Román CSIC ORCID; Cabello-Lobato, María J. CSIC; Prado, Félix CSIC ORCID
Palabras claveChromatin endogenous cleavage
DNA binding proteins
Non-double-strand break
Rad52
Replication intermediates
Rad51
Replication fork
2D DNA gel electrophoresis
Fecha de publicación2021
EditorHumana Press
Springer Nature
CitaciónHomologous Recombination. Methods and Protocols: 447-458 (2021)
SerieMethods in Molecular Biology
2153
ResumenHomologous 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.
Versión del editorhttps://doi.org/10.1007/978-1-0716-0644-5_31
URIhttp://hdl.handle.net/10261/250707
DOI10.1007/978-1-0716-0644-5_31
ISBN978-1-0716-0643-8
ISSN1064-3745
E-ISSN1940-6029
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