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Molecular and Cellular Biology, May 2007, p. 3828-3838, Vol. 27, No. 10
0270-7306/07/$08.00+0     doi:10.1128/MCB.01596-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Mus81-Eme1-Dependent and -Independent Crossovers Form in Mitotic Cells during Double-Strand Break Repair in Schizosaccharomyces pombe{triangledown}

Justin C. Hope,1 Lissette Delgado Cruzata,2 Amit Duvshani,3 Jun Mitsumoto,2 Mohamed Maftahi,2 and Greg A. Freyer1,2*

Graduate Program in Anatomy and Cell Biology,1 Graduate Program in Environmental Health Sciences,2 Graduate Program in Physiology and Cellular Biophysics, Columbia University, 722 West 168th St., Kolb Bldg., Rm. 140, New York, New York 100323

Received 26 August 2006/ Returned for modification 18 September 2006/ Accepted 2 March 2007

During meiosis, double-strand breaks (DSBs) lead to crossovers, thought to arise from the resolution of double Holliday junctions (HJs) by an HJ resolvase. In Schizosaccharomyces pombe, meiotic crossovers are produced primarily through a mechanism requiring the Mus81-Eme1 endonuclease complex. Less is known about the processes that produces crossovers during the repair of DSBs in mitotic cells. We employed an inducible DSB system to determine the role of Rqh1-Top3 and Mus81-Eme1 in mitotic DSB repair and crossover formation in S. pombe. In agreement with the meiotic data, crossovers are suppressed in cells lacking Mus81-Eme1. And relative to the wild type, rqh1{Delta} cells show a fourfold increase in crossover frequency. This suppression of crossover formation by Rqh1 is dependent on its helicase activity. We found that the synthetic lethality of cells lacking both Rqh1 and Eme1 is suppressed by loss of swi5+, which allowed us to show that the excess crossovers formed in an rqh1{Delta} background are independent of Mus81-Eme1. This result suggests that a second process for crossover formation exists in S. pombe and is consistent with our finding that deletion of swi5+ restored meiotic crossovers in eme1{Delta} cells. Evidence suggesting that Rqh1 also acts downstream of Swi5 in crossover formation was uncovered in these studies. Our results suggest that during Rhp51-dependent repair of DSBs, Rqh1-Top3 suppresses crossovers in the Rhp57-dependent pathway while Mus81-Eme1 and possibly Rqh1 promote crossovers in the Swi5-dependent pathway.

* Corresponding author. Mailing address: 722 W. 168th St., Kolb Bldg., Rm. 140, New York, NY 10032. Phone: (212) 342-0457. Fax: (212) 781-4993. E-mail: gaf1{at}columbia.edu

{triangledown} Published ahead of print on 12 March 2007.

Molecular and Cellular Biology, May 2007, p. 3828-3838, Vol. 27, No. 10
0270-7306/07/$08.00+0     doi:10.1128/MCB.01596-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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