Functional Centromeres Determine the Activation Time of Pericentric Origins of DNA Replication in Saccharomyces cerevisiae 英文参考文献.docVIP

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Functional Centromeres Determine the Activation Time of Pericentric Origins of DNA Replication in Saccharomyces cerevisiae 英文参考文献.doc

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Functional Centromeres Determine the Activation Time of Pericentric Origins of DNA Replication in Saccharomyces cerevisiae 英文参考文献

FunctionalCentromeresDeterminetheActivationTime ofPericentricOriginsofDNAReplicationin Saccharomycescerevisiae ThomasJ.Pohl1,2,BonitaJ.Brewer1,2,M.K.Raghuraman2* 1Molecular and Cellular Biology Program, University of Washington, Seattle, Washington, United States of America, 2Department of Genome Sciences, University of Washington,Seattle,Washington,UnitedStatesofAmerica Abstract ThecentromericregionsofallSaccharomycescerevisiaechromosomesarefoundinearlyreplicatingdomains,aproperty conserved among centromeres in fungi and some higher eukaryotes. Surprisingly, little is known about the biological significance or the mechanism of early centromere replication; however, the extensive conservation suggests that it is importantfor chromosomemaintenance.Do centromeres ensuretheir earlyreplication bypromoting earlyactivation of nearbyorigins,orhavetheymigratedoverevolutionarytimetoresideinearlyreplicatingregions?InCandidaalbicans ,a neocentromerecontainsanearlyfiringorigin,supportingthefirsthypothesisbutnotaddressingwhethertheneworiginis intrinsically early firing or whether the centromere influences replication time. Because the activation time of individual originsisnotanintrinsicpropertyofS.cerevisiaeorigins,butisinfluencedbysurroundingsequences,wesoughttotestthe hypothesisthatcentromeresinfluencereplicationtimebymovingacentromeretoalatereplicationdomain.Weuseda modified Meselson-Stahl density transfer assay to measure the kinetics of replication for regions of chromosome XIV in which either the functional centromere or a point-mutated version had been moved near origins that reside in a late replication region. We show that a functional centromere acts in cis over a distance as great as 19kb to advance the initiationtimeoforigins.Ourresultsconstituteadirectlinkbetweenestablishmentofthekinetochoreandthereplication initiation machinery, and suggest that the proposed higher-order structure of the pericentric chromatin influences replicationinitiation. Citation: Pohl TJ,