Advertisement

Methods to Map Meiotic Recombination Proteins in Saccharomyces cerevisiae

  • Aurore Sanchez
  • Valérie BordeEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2153)

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.

Key words

Chromatin immunoprecipitation High-throughput sequencing Genome-wide maps DNA sonication Meiotic proteins Recombination intermediate mapping 

References

  1. 1.
    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.2011.02.009CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Hunter N (2015) Meiotic recombination: the essence of heredity. Cold Spring Harb Perspect Biol.  https://doi.org/10.1101/cshperspect.a016618
  3. 3.
    Muller HJ (1916) The mechanism of crossing over. Am Nat 50:193–434CrossRefGoogle Scholar
  4. 4.
    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.2008.07.006CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    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.2012.03.025CrossRefPubMedGoogle Scholar
  6. 6.
    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.1003416CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    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
  8. 8.
    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.308510.117CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    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 557:117–142CrossRefGoogle Scholar
  10. 10.
    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):59–70CrossRefGoogle Scholar
  11. 11.
    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.0125965CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  1. 1.Institut Curie – Research Center, UMR3244 CNRS, Pavillon Trouillet RossignolPSL Research UniversityParis Cedex 05France
  2. 2.Paris Sorbonne UniversitéParisFrance

Personalised recommendations