Genome-Wide Redistribution of Meiotic Double-Strand Breaks in Saccharomyces cerevisiae

Abstract

Meiotic recombination is initiated by the formation of programmed DNA double-strand breaks (DSBs) catalyzed by the Spo11 protein. DSBs are not randomly distributed along chromosomes. To better understand factors that control the distribution of DSBs in budding yeast, we have examined the genome-wide binding and cleavage properties of the Gal4 DNA binding domain (Gal4BD)-Spo11 fusion protein. We found that Gal4BD-Spo11 cleaves only a subset of its binding sites, indicating that the association of Spo11 with chromatin is not sufficient for DSB formation. In centromere-associated regions, the centromere itself prevents DSB cleavage by tethered Gal4BD-Spo11 since its displacement restores targeted DSB formation. In addition, we observed that new DSBs introduced by Gal4BD-Spo11 inhibit surrounding DSB formation over long distances (up to 60 kb), keeping constant the number of DSBs per chromosomal region. Together, these results demonstrate that the targeting of Spo11 to new chromosomal locations leads to both local stimulation and genome-wide redistribution of recombination initiation and that some chromosomal regions are inherently cold regardless of the presence of Spo11.

SUPPLEMENTAL MATERIAL

We thank Tzu-Chen Wu and Michael Lichten for providing the cen3Δ strain and communicating results before publication and Richard Durbin from the Sanger Institute and Ed Louis for allowing us to use the unpublished SK1 sequence information. We thank Michèle Vedel for constructing the pAP11 plasmid. We also thank Christine Mézard and Pierre-Antoine Defossez for comments that improved the manuscript and the anonymous reviewers for helpful suggestions.

This work was supported by grants from the Association pour la Recherche sur le Cancer (to A.N.). N.R. was supported by a predoctoral fellowship from Institut Curie. N.U. was supported by postdoctoral fellowships from the CNRS and the Institut Curie.