Cdk1 Targets Srs2 to Complete Synthesis-Dependent Strand Annealing and to Promote Recombinational Repair 英文参考文献.docVIP

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Cdk1 Targets Srs2 to Complete Synthesis-Dependent Strand Annealing and to Promote Recombinational Repair 英文参考文献.doc

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Cdk1 Targets Srs2 to Complete Synthesis-Dependent Strand Annealing and to Promote Recombinational Repair 英文参考文献

Cdk1TargetsSrs2toCompleteSynthesis-Dependent StrandAnnealingandtoPromoteRecombinational Repair MarcoSaponaro1¤a,DevonCallahan2¤b,XiuzhongZheng2,LumirKrejci3,JamesE.Haber4,HannahL. Klein2,GiordanoLiberi1* 1InstituteofMolecularOncologyFoundation,TheItalianFoundationforCancerResearchandDipartimentodiScienzeBiomolecolarieBiotecnologie–UniversityofMilan, Milan, Italy, 2Department of Biochemistry and Kaplan Cancer Center, New York University School of Medicine, New York, New York, United States of America, 3Department of Biology and National Center for Biomolecular Research, Masaryk University, Brno, Czech Republic, 4Department of Biology and Rosenstiel Medical Center,BrandeisUniversity,Waltham,Massachusetts,UnitedStatesofAmerica Abstract Cdk1kinasephosphorylates buddingyeastSrs2, amemberof UvrDproteinfamily,displays bothDNA translocation and DNA unwinding activities in vitro. Srs2 prevents homologous recombination by dismantling Rad51 filaments and is also required for double-strand break (DSB) repair. Here we examine the biological significance of Cdk1-dependent phosphorylation of Srs2, using mutants that constitutively express the phosphorylated or unphosphorylated protein isoforms. We found that Cdk1 targets Srs2 to repair DSB and, in particular, to complete synthesis-dependent strand annealing,likelycontrollingthedisassemblyofaD-loopintermediate.Cdk1-dependentphosphorylationcontrolsturnover ofSrs2attheinvadingstrand;and,inabsenceofthismodification,theturnoverofRad51isnotaffected.Furtheranalysisof the recombination phenotypes of the srs2 phospho-mutants showed that Srs2 phosphorylation is not required for the removal of toxic Rad51 nucleofilaments, although it is essential for cell survival, when DNA breaks are channeled into homologous recombinational repair. Cdk1-targeted Srs2 displays a PCNA–independent role and appears to have an attenuatedabilitytoinhibitrecombination.Finally,therecombinationdefectsofunphosphorylatableSrs2areprimarilydue to unscheduled acc