In vivo binding of recombination proteins to non-DSB DNA lesions and to replication forks
Affiliation
Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, The Netherlands.Issue Date
2021
Metadata
Show full item recordAbstract
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.Citation
Gonzalez-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.Journal
Methods in Molecular BiologyDOI
10.1007/978-1-0716-0644-5_31PubMed ID
32840798Additional Links
https://dx.doi.org/10.1007/978-1-0716-0644-5_31Type
ArticleLanguage
enae974a485f413a2113503eed53cd6c53
10.1007/978-1-0716-0644-5_31
Scopus Count
Collections
Related articles
- Rad51 replication fork recruitment is required for DNA damage tolerance.
- Authors: González-Prieto R, Muñoz-Cabello AM, Cabello-Lobato MJ, Prado F
- Issue date: 2013 May 2
- Differential regulation of homologous recombination at DNA breaks and replication forks by the Mrc1 branch of the S-phase checkpoint.
- Authors: Alabert C, Bianco JN, Pasero P
- Issue date: 2009 Apr 22
- Ctf4 Prevents Genome Rearrangements by Suppressing DNA Double-Strand Break Formation and Its End Resection at Arrested Replication Forks.
- Authors: Sasaki M, Kobayashi T
- Issue date: 2017 May 18
- A postincision-deficient TFIIH causes replication fork breakage and uncovers alternative Rad51- or Pol32-mediated restart mechanisms.
- Authors: Moriel-Carretero M, Aguilera A
- Issue date: 2010 Mar 12
- The role of DNA double-strand breaks in spontaneous homologous recombination in S. cerevisiae.
- Authors: Lettier G, Feng Q, de Mayolo AA, Erdeniz N, Reid RJ, Lisby M, Mortensen UH, Rothstein R
- Issue date: 2006 Nov 10