Sgs1 and Exo1 Redundantly Inhibit Break-Induced Replication and De Novo Telomere Addition at Broken Chromosome Ends 英文参考文献.docVIP

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Sgs1 and Exo1 Redundantly Inhibit Break-Induced Replication and De Novo Telomere Addition at Broken Chromosome Ends 英文参考文献.doc

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Sgs1 and Exo1 Redundantly Inhibit Break-Induced Replication and De Novo Telomere Addition at Broken Chromosome Ends 英文参考文献

Sgs1andExo1RedundantlyInhibitBreak-Induced ReplicationandDeNovoTelomereAdditionatBroken ChromosomeEnds JohnR.Lydeard¤,ZacharyLipkin-Moore,SuviJain,VinayV.Eapen,JamesE.Haber* DepartmentofBiologyandRosenstielBasicMedicalSciencesResearchCenter,BrandeisUniversity,Waltham,Massachusetts,UnitedStatesofAmerica Abstract Inbuddingyeast,anHOendonuclease-inducible double-strand break(DSB)isefficientlyrepairedbyseveralhomologous recombination (HR)pathways.Incontrasttogeneconversion (GC),wherebothendsoftheDSBcanrecombinewiththe same template, break-induced replication (BIR) occurs when only the centromere-proximal end of the DSB can locate homologous sequences. Whereas GC results in a small patch of new DNA synthesis, BIR leads to a nonreciprocal translocation. The requirements for completing BIR are significantly different from those of GC, but both processes require59to39resectionofDSBendstocreatesingle-stranded DNAthatleadstoformationofaRad51filamentrequired toinitiate HR.Resection proceedsbytwopathwaysdependentonExo1ortheBLMhomolog,Sgs1.WereportthatExo1 and Sgs1 each inhibit BIR but have little effect on GC, while overexpression of either protein severely inhibits BIR. In contrast, overexpression ofRad51markedly increases theefficiency ofBIR,againwithlittleeffectonGC.Insgs1Dexo1D strains,wherethereislittle59to39resection,thelevelofBIRisnotdifferentfromeithersinglemutant;surprisingly, there isatwo-foldincreaseincellviability afterHOinductionwhereby40%ofallcellssurvivebyformation ofanewtelomere withinafewkbofthesiteofDNAcleavage.Denovotelomereadditionisrareinwild-type,sgs1D,orexo1Dcells.Insgs1D exo1D,repairbyGCisseverely inhibited, butcellviaiblity remains highbecause ofnewtelomere formation. Thesedata suggest thattheextensive 59to39resection thatoccurs before theinitiation ofnewDNAsynthesis inBIRmayprevent efficient maintenance of a Rad51 filament near the DSB end. The severe constraint on 59 to 39 resection, which also abrogatesactivation ofth