Abstract
Homologous 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.
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Acknowledgments
This work was supported by grants BFU2012-38171 and BFU2015-63698-P from the Spanish government.
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González-Prieto, R., Cabello-Lobato, M.J., Prado, F. (2021). In Vivo Binding of Recombination Proteins to Non-DSB DNA Lesions and to Replication Forks. In: Aguilera, A., Carreira, A. (eds) Homologous Recombination. Methods in Molecular Biology, vol 2153. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0644-5_31
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DOI: https://doi.org/10.1007/978-1-0716-0644-5_31
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