Sensing of Replication Stress and Mec1 Activation Act through Two Independent Pathways Involving the 9-1-1 Complex and DNA Polymerase ε 英文参考文献.docVIP

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Sensing of Replication Stress and Mec1 Activation Act through Two Independent Pathways Involving the 9-1-1 Complex and DNA Polymerase ε 英文参考文献.doc

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Sensing of Replication Stress and Mec1 Activation Act through Two Independent Pathways Involving the 9-1-1 Complex and DNA Polymerase ε 英文参考文献

SensingofReplicationStressandMec1ActivationAct throughTwoIndependentPathwaysInvolvingthe9-1-1 ComplexandDNAPolymerasee FabioPuddu,GabrielePiergiovanni¤,PaoloPlevani*,MarcoMuzi-Falconi* DepartmentofBiomolecularSciencesandBiotechnology,Universita` degliStudidiMilano,Milano,Italy Abstract FollowingDNAdamageorreplicationstress,buddingyeastcellsactivatetheRad53checkpointkinase,promotinggenome stabilityinthesechallengingconditions.TheDNAdamageandreplicationcheckpointpathwaysarepartiallyoverlapping, sharingseveralfactors,butarealsodifferentiatedatvariouslevels.TheupstreamkinaseMec1isrequiredtoactivateboth signalingcascadestogetherwiththe9-1-1PCNA-likecomplexandtheDpb11(hTopBP1)protein.AfterDNAdamage,Dpb11is alsoneededtorecruittheadaptorproteinRad9(h53BP1).HereweanalyzedthemechanismsleadingtoMec1activationinvivo afterDNAdamageandreplicationstress.Wefoundthataddc1Ddpb11-1doublemutantstraindisplaysasyntheticdefectin Rad53andH2Aphosphorylationandisextremelysensitivetohydroxyurea(HU),indicatingthatDpb11andthe9-1-1complex independentlypromoteMec1activation.Asimilarphenotypeisobservedwhenboththe9-1-1complexandtheDpb4non- essentialsubunitofDNApolymerasee(Pole)arecontemporarilyabsent,indicatingthatcheckpointactivationinresponseto replicationstressisachievedthroughtwoindependentpathways,requiringthe9-1-1complexandPole. Citation:PudduF,PiergiovanniG,PlevaniP,Muzi-FalconiM(2011)SensingofReplicationStressandMec1ActivationActthroughTwoIndependentPathways Involvingthe9-1-1ComplexandDNAPolymerasee.PLoSGenet7(3):e1002022.doi:10.1371/journal.pgen.1002022 Editor:GregoryP.Copenhaver,TheUniversityofNorthCarolinaatChapelHill,UnitedStatesofAmerica ReceivedNovember10,2010;AcceptedJanuary20,2011;PublishedMarch17,2011 Copyright: ? 2011 Puddu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:Thisworkwassupported