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Molecular and Cellular Biology, March 2007, p. 1868-1880, Vol. 27, No. 5
0270-7306/07/$08.00+0     doi:10.1128/MCB.02063-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Genome-Wide Redistribution of Meiotic Double-Strand Breaks in Saccharomyces cerevisiae{triangledown} ,{dagger}

Nicolas Robine,1,2 Norio Uematsu,1,{ddagger} Franck Amiot,3 Xavier Gidrol,3 Emmanuel Barillot,2 Alain Nicolas,1 and Valérie Borde1*

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,1 Institut Curie, Service de Bioinformatique, 26 rue d'Ulm, 75248 Paris Cedex 05, France,2 and Service de Génomique Fonctionnelle, CEA, 2 rue Gaston Crémieux, CP5722, 91057 Evry Cedex, France3

Received 3 November 2006/ Returned for modification 30 November 2006/ Accepted 12 December 2006

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.

* Corresponding author. Mailing address: 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. Phone: 33 (0) 1 42 34 66 37. Fax: 33 (0) 1 42 34 66 44. E-mail: valerie.borde{at}curie.fr.

{triangledown} Published ahead of print on 22 December 2006.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

{ddagger} Present address: Kyoto University, Radiation Biology Center, Late Effect Studies Konoe-cho, Yoshida, Sakyo-ku, Kyoto-shi 606-8501, Japan.

Molecular and Cellular Biology, March 2007, p. 1868-1880, Vol. 27, No. 5
0270-7306/07/$08.00+0     doi:10.1128/MCB.02063-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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