Distinct Cdk1 Requirements during Single-Strand Annealing, Noncrossover, and Crossover Recombination 英文参考文献.docVIP

DistinctCdk1RequirementsduringSingle-Strand Annealing,Noncrossover,andCrossoverRecombination CamillaTrovesi,MarcoFalcettoni,GiovannaLucchini,MichelaClerici*,MariaPiaLonghese* DipartimentodiBiotecnologieeBioscienze,Universita` diMilano-Bicocca,Milano,Italy Abstract RepairofDNAdouble-strandbreaks(DSBs)byhomologousrecombination(HR)inhaploidcellsisgenerallyrestrictedtoS/ G2cellcyclephases,whenDNAhasbeenreplicatedandasisterchromatidisavailableasarepairtemplate.Thiscellcycle specificitydependsoncyclin-dependentproteinkinases(Cdk1inSaccharomycescerevisiae),whichinitiateHRbypromoting 59–39 nucleolytic degradation of the DSB ends. Whether Cdk1 regulates other HR steps is unknown. Here we show that yku70Dcells,whichaccumulatesingle-strandedDNA(ssDNA)attheDSBendsindependentlyofCdk1activity,areableto repairaDSBbysingle-strandannealing(SSA)intheG1cellcyclephase,whenCdk1activityislow.Thisabilitytoperform SSA depends on DSB resection, because both resection and SSA are enhanced by the lack of Rad9 in yku70D G1 cells. Furthermore,wefoundthatinterchromosomalnoncrossoverrecombinantsaregeneratedinyku70Dandyku70Drad9DG1 cells, indicating that DSB resection bypasses Cdk1 requirement also for carrying out these recombination events. By contrast,yku70Dandyku70Drad9DcellsarespecificallydefectiveininterchromosomalcrossoverrecombinationwhenCdk1 activity is low. Thus, Cdk1 promotes DSB repair by single-strand annealing and noncrossover recombination by acting mostlyattheresectionlevel,whereasadditionaleventsrequireCdk1-dependentregulationinordertogeneratecrossover outcomes. Citation: Trovesi C, Falcettoni M, Lucchini G, Clerici M, Longhese MP (2011) Distinct Cdk1 Requirements during Single-Strand Annealing, Noncrossover, and CrossoverRecombination.PLoSGenet7(8):e1002263.doi:10.1371/journal.pgen.1002263 Editor:SueBiggins,FredHutchinsonCancerResearchCenter,UnitedStatesofAmerica ReceivedApril28,2011;AcceptedJuly8,2011;PublishedAugust25,2011 Copyright: ? 2011 Trovesi et al. This is an open-ac