Genome-wide redistribution of meiotic double-strand breaks in Saccharomyces cerevisiae.
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.
Robine
Nicolas
N
Institut Curie, Recombinaison et Instabilité Génétique, Centre de Recherche, UMR7147 CNRS-Institut Curie-Université P. et M. Curie, 26 rue d'Ulm, 75248 Paris Cedex 05, France. valerie.borde@curie.fr
Uematsu
Norio
N
Amiot
Franck
F
Gidrol
Xavier
X
Barillot
Emmanuel
E
Nicolas
Alain
A
Borde
Valérie
V
D001665:Binding Sites; D047369:Chromatin Immunoprecipitation; D053903:DNA Breaks, Double-Stranded; D004271:DNA, Fungal / Q000235:genetics*; D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases; D004950:Esterases / Q000235:genetics / Q000378:metabolism; D016681:Genome, Fungal; D008540:Meiosis; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D000099257:Meiotic Recombination Protein SPO11
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases; D004950:Esterases; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D000099257:Meiotic Recombination Protein SPO11; C052130:GAL4 protein, S cerevisiae