A Dynamic Stochastic Model for DNA Replication Initiation in Early Embryos 英文参考文献.docVIP

ADynamicStochasticModelforDNAReplication InitiationinEarlyEmbryos ArachGoldar1*,He′le`neLabit2¤,KathrinMarheineke2,OlivierHyrien2* 1ServicedeBiologieInte′grative etdeGe′ne′tiqueMole′culaire, Commissariata` l’E′nergieAtomique,Gif-sur-Yvette,France,2EcoleNormaleSupe′rieure,CNRSUMR8541, Paris,France Abstract Background: Eukaryotic cells seem unable to monitor replication completion during normal S phase, yet must ensure a reliablereplicationcompletiontime.ThisisanacuteprobleminearlyXenopusembryossinceDNAreplicationoriginsare located and activated stochastically, leading to the random completion problem. DNA combing, kinetic modelling and other studies using Xenopus egg extracts have suggested that potential origins are much more abundant than actual initiationeventsandthatthetime-dependentrateofinitiation,I(t),markedlyincreasesthroughSphasetoensuretherapid completionofunreplicatedgapsandanarrowdistributionofcompletiontimes.However,themolecularmechanismthat underliesthisincreasehasremainedobscure. Methodology/PrincipalFindings:UsingbothpreviousandnovelDNAcombingdatawehaveconfirmedthatI(t)increases throughSphasebuthavealsoestablishedthatitprogressivelydecreasesbeforetheendofSphase.Toexploreplausible biochemicalscenariosthatmightexplainthesefeatures,wehaveperformedcomparisonsbetweennumericalsimulations and DNA combing data. Several simple models were tested: i) recycling of a limiting replication fork component from completedreplicons;ii)time-dependentincreaseinoriginefficiency;iii)time-dependentincreaseinavailabilityofaninitially limiting factor, e.g. by nuclear import. None of these potential mechanisms could on its own account for the data. We proposeamodelthatcombinestime-dependentchangesinavailabilityofareplicationfactorandafork-densitydependent affinityofthisfactorforpotentialorigins.Thisnovelmodelquantitativelyandrobustlyaccountedfortheobservedchanges ininitiationrateandforkdensity. Conclusions/Significance:Thisworkprovidesarefinedtemporalprofileofreplicationi