Abstract
Meiotic recombination is triggered by programmed DNA double-strand breaks (DSBs), catalyzed by the type II topoisomerase-like Spo11 protein. Meiotic DSBs are repaired by homologous recombination, which produces either crossovers or noncrossovers, this decision being linked to the binding of proteins specific of each pathway. Mapping the binding of these proteins along chromosomes in wild type or mutant yeast background is extremely useful to understand how and at which step the decision to repair a DSB with a crossover is taken. It is now possible to obtain highly synchronous yeast meiotic populations, which, combined with appropriate negative controls, enable to detect by chromatin immunoprecipitation followed by sequencing (ChIP-Seq) the transient binding of diverse recombination proteins with high sensitivity and resolution.
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References
Pan J, Sasaki M, Kniewel R, Murakami H, Blitzblau HG, Tischfield SE, Zhu X, Neale MJ, Jasin M, Socci ND, Hochwagen A, Keeney S (2011) A hierarchical combination of factors shapes the genome-wide topography of yeast meiotic recombination initiation. Cell 144(5):719–731. https://doi.org/10.1016/j.cell.2011.02.009
Hunter N (2015) Meiotic recombination: the essence of heredity. Cold Spring Harb Perspect Biol. https://doi.org/10.1101/cshperspect.a016618
Muller HJ (1916) The mechanism of crossing over. Am Nat 50:193–434
Chen SY, Tsubouchi T, Rockmill B, Sandler JS, Richards DR, Vader G, Hochwagen A, Roeder GS, Fung JC (2008) Global analysis of the meiotic crossover landscape. Dev Cell 15(3):401–415. https://doi.org/10.1016/j.devcel.2008.07.006
Serrentino ME, Borde V (2012) The spatial regulation of meiotic recombination hotspots: are all DSB hotspots crossover hotspots? Exp Cell Res 318(12):1347–1352. https://doi.org/10.1016/j.yexcr.2012.03.025
Serrentino ME, Chaplais E, Sommermeyer V, Borde V (2013) Differential association of the conserved SUMO ligase Zip3 with meiotic double-strand break sites reveals regional variations in the outcome of meiotic recombination. PLoS Genet 9(4):e1003416. https://doi.org/10.1371/journal.pgen.1003416
Chia M, van Werven FJ (2016) Temporal expression of a master regulator drives synchronous sporulation in budding yeast. G3 (Bethesda). https://doi.org/10.1534/g3.116.034983
De Muyt A, Pyatnitskaya A, Andreani J, Ranjha L, Ramus C, Laureau R, Fernandez-Vega A, Holoch D, Girard E, Govin J, Margueron R, Coute Y, Cejka P, Guerois R, Borde V (2018) A meiotic XPF-ERCC1-like complex recognizes joint molecule recombination intermediates to promote crossover formation. Genes Dev 32(3-4):283–296. https://doi.org/10.1101/gad.308510.117
Murakami H, Borde V, Nicolas A, Keeney S (2009) Gel electrophoresis assays for analyzing DNA double-strand breaks in Saccharomyces cerevisiae at various spatial resolutions. Methods Mol Biol 557:117–142
Hunter N, Kleckner N (2001) The single-end invasion: an asymmetric intermediate at the double-strand break to double-holliday junction transition of meiotic recombination. Cell 106(1):59–70
Brachet E, Beneut C, Serrentino ME, Borde V (2015) The CAF-1 and Hir histone chaperones associate with sites of meiotic double-strand breaks in budding yeast. PLoS One 10:e0125965. https://doi.org/10.1371/journal.pone.0125965
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Sanchez, A., Borde, V. (2021). Methods to Map Meiotic Recombination Proteins in Saccharomyces cerevisiae. 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_21
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DOI: https://doi.org/10.1007/978-1-0716-0644-5_21
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