Mus81-Eme1-dependent and -independent crossovers form in mitotic cells during double-strand break repair in Schizosaccharomyces pombe.
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, rqh1Delta 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 rqh1Delta 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 eme1Delta 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.
Hope
Justin C
JC
Graduate Program in Anatomy and Cell Biology, Columbia University, 722 W. 168th Street, New York, NY 10032, USA.
Cruzata
Lissette Delgado
LD
Duvshani
Amit
A
Mitsumoto
Jun
J
Maftahi
Mohamed
M
Freyer
Greg A
GA
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D003434:Crossing Over, Genetic; D053903:DNA Breaks, Double-Stranded; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair; D004264:DNA Topoisomerases, Type I / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D016521:Electrophoresis, Gel, Pulsed-Field; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism*; D005785:Gene Conversion; D008938:Mitosis / Q000502:physiology*; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000166:cytology / Q000235:genetics / Q000502:physiology; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*
D004268:DNA-Binding Proteins; C495865:MUS81 protein, S pombe; C084232:RHP51 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; C483769:Swi5 protein, S pombe; D051135:Rad51 Recombinase; C442837:Eme1 protein, S pombe; D004720:Endonucleases; D000251:Adenosine Triphosphatases; C442472:Rhp57 protein, S pombe; D004265:DNA Helicases; C106462:Rqh1 protein, S pombe; D004264:DNA Topoisomerases, Type I