Preferential Localization of Human Origins of DNA Replication at the 5′-Ends of Expressed Genes and at Evolutionarily Conserved DNA Sequences 英文参考文献.docVIP

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Preferential Localization of Human Origins of DNA Replication at the 5′-Ends of Expressed Genes and at Evolutionarily Conserved DNA Sequences 英文参考文献.doc

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Preferential Localization of Human Origins of DNA Replication at the 5′-Ends of Expressed Genes and at Evolutionarily Conserved DNA Sequences 英文参考文献

PreferentialLocalizationofHumanOriginsofDNA Replicationatthe59-EndsofExpressedGenesandat EvolutionarilyConservedDNASequences ManuelS.Valenzuela1,2*.,YidongChen1.¤,SeanDavis1,FanYang1,RobertL.Walker1,SvenBilke1 ,John Lueders1,MelveniaM.Martin3,MiritI.Aladjem3,PierreP.Massion4,PaulS.Meltzer1* 1GeneticsBranch,CenterforCancerResearch,NationalCancerInstitute,NationalInstitutesofHealth,Bethesda,Maryland,UnitedStatesofAmerica,2Departmentof BiochemistryandCancerBiology,MeharryMedicalCollege,Nashville,Tennessee,UnitedStatesofAmerica,3LaboratoryofMolecularPharmacology,CenterforCancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of America, 4Division of Allergy, Pulmonary and Critical Care Medicine,VanderbiltIngramCancerCenter,VanderbiltUniversity,Nashville,Tennessee,UnitedStatesofAmerica Abstract Background:Replicationofmammaliangenomesrequirestheactivationofthousandsoforiginswhicharebothspatially and temporally regulated by as yet unknown mechanisms. At the most fundamental level, our knowledge about the distributionpatternoforiginsineachofthechromosomes,amongdifferentcelltypes,andwhetherthephysiologicalstate ofthecellsaltersthisdistributionisatpresentverylimited. Methodology/PrincipalFindings:Wehaveusedstandardl-exonucleaseresistantnascentDNApreparationsinthesizerange of0.7–1.5kbobtainedfromthebreastcancercelllineMCF–7hybridizedtoacustomtilingarraycontaining50–60ntprobes evenlydistributedamonggenicandnon-genicregionscoveringabout1%ofthehumangenome.AsimilarDNApreparation wasusedforhigh-throughputDNAsequencing.ArrayexperimentswerealsoperformedwithDNAobtainedfromBT-474and H520celllines.BydeterminingthesitesshowingnascentDNAenrichment,wehavelocalizedseveralthousandoriginsofDNA replication.Ourmajorfindingsare:(a)botharrayandDNAsequencingassaymethodsproducedessentiallythesameorigin distributionprofile;(b)origindistributionislargelyconserved(.70%)inallcelllinestested;(c)originsareenrichedatthe 59endsofexpressedgenesandatev