Abstract
Homologous recombination is a high-fidelity DNA double-strand break repair pathway that uses a homologous template to repair the break. Recombinases are the central enzymes that facilitate the strand invasion step of homologous recombination, which forms a DNA joint molecule. These DNA joint molecules can be moved through branch migration activity. In this chapter, we describe two assays to determine the branch migration activity and directionality of an enzyme. Monitoring the branch migration activity of an enzyme can provide insight into the roles of these factors in homologous recombination.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
San Filippo J, Sung P, Klein H (2008) Mechanism of eukaryotic homologous recombination. Annu Rev Biochem 77:229–257
Bugreev DV, Hanaoka F, Mazin AV (2007) Rad54 dissociates homologous recombination intermediates by branch migration. Nat Struct Mol Biol 14:746–753
Sugiyama T, Kantake N, Wu Y, Kowalczykowski SC (2006) Rad52-mediated DNA annealing after Rad51-mediated DNA strand exchange promotes second ssDNA capture. EMBO J 25:5539–5548
Holliday R (1964) A mechanism for gene conversion in fungi. Genet Res 5:282–304
Paques F, Haber JE (1999) Multiple pathways of recombination induced by double-strand breaks in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 63:349–404
Wyatt HD, West SC (2014) Holliday junction resolvases. Cold Spring Harb Perspect Biol 6:a023192
Seigneur M, Bidnenko V, Ehrlich SD, Michel B (1998) RuvAB acts at arrested replication forks. Cell 95:419–430
McGlynn P, Lloyd RG, Marians KJ (2001) Formation of Holliday junctions by regression of nascent DNA in intermediates containing stalled replication forks: RecG stimulates regression even when the DNA is negatively supercoiled. Proc Natl Acad Sci U S A 98:8235–8240
Betous R, Mason AC, Rambo RP, Bansbach CE, Badu-Nkansah A, Sirbu BM, Eichman BF, Cortez D (2012) SMARCAL1 catalyzes fork regression and Holliday junction migration to maintain genome stability during DNA replication. Genes Dev 26:151–162
Bugreev DV, Mazina OM, Mazin AV (2006) Rad54 protein promotes branch migration of Holliday junctions. Nature 442:590–593
Pike AC, Gomathinayagam S, Swuec P, Berti M, Zhang Y, Schnecke C, Marino F, von Delft F, Renault L, Costa A et al (2015) Human RECQ1 helicase-driven DNA unwinding, annealing, and branch migration: insights from DNA complex structures. Proc Natl Acad Sci U S A 112:4286–4291
Bizard AH, Hickson ID (2014) The dissolution of double Holliday junctions. Cold Spring Harb Perspect Biol 6:a016477
Chi P, Van Komen S, Sehorn MG, Sigurdsson S, Sung P (2006) Roles of ATP binding and ATP hydrolysis in human Rad51 recombinase function. DNA Repair 5:381–391
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Kelso, A.A., Goodson, S.D., Sehorn, M.G. (2019). In Vitro Assays for DNA Branch Migration. In: Balakrishnan, L., Stewart, J. (eds) DNA Repair. Methods in Molecular Biology, vol 1999. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9500-4_18
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9500-4_18
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9499-1
Online ISBN: 978-1-4939-9500-4
eBook Packages: Springer Protocols