Methods to Map Meiotic Recombination Proteins in Saccharomyces cerevisiae

Methods Mol Biol. 2021:2153:295-306. doi: 10.1007/978-1-0716-0644-5_21.

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.

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

MeSH terms

  • Chromatin Immunoprecipitation Sequencing
  • Chromosome Mapping / methods*
  • Endodeoxyribonucleases / genetics*
  • High-Throughput Nucleotide Sequencing
  • Meiosis
  • Mutation
  • Recombinational DNA Repair
  • Saccharomyces cerevisiae / genetics*

Substances

  • Endodeoxyribonucleases
  • meiotic recombination protein SPO11