Association of the Chromosome Replication Initiator DnaA with the Escherichia coli Inner Membrane In Vivo Quantity and Mode of Binding 英文参考文献.docVIP

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Association of the Chromosome Replication Initiator DnaA with the Escherichia coli Inner Membrane In Vivo Quantity and Mode of Binding 英文参考文献.doc

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Association of the Chromosome Replication Initiator DnaA with the Escherichia coli Inner Membrane In Vivo Quantity and Mode of Binding 英文参考文献

AssociationoftheChromosomeReplicationInitiator DnaAwiththeEscherichiacoliInnerMembraneInVivo: QuantityandModeofBinding TomerRegev1,NadavMyers1,RazZarivach1,2,ItzhakFishov1* 1DepartmentofLifeSciences,BenGurionUniversityoftheNegev,Beer-Sheva,Israel,2NationalInstituteofBiotechnology,Ben-GurionUniversityoftheNegev,Beer- Sheva,Israel Abstract DnaAinitiateschromosomereplicationinmostknownbacteriaanditsactivityiscontrolledsothatthiseventoccursonly onceeverycelldivisioncycle.ATPintheactiveATP-DnaAishydrolyzedafterinitiationandtheresultingADPisreplaced withATPonthevergeofthenextinitiation.TwoputativerecyclingmechanismsdependonthebindingofDnaAeitherto the membrane or to specific chromosomal sites, promoting nucleotide dissociation. While there is no doubt that DnaA interactswithartificialmembranesinvitro,itisstillcontroversialastowhetheritbindsthecytoplasmicmembraneinvivo .In thisworkwelookedforDnaA-membraneinteractioninE.colicellsbyemployingcellfractionationwithbothnativeand fluorescent DnaA hybrids. We show that about 10% of cellular DnaA is reproducibly membrane-associated. This small fraction might be physiologically significant and represent the free DnaA available for initiation, rather than the vast majorityboundtothedatAreservoir.UsingthecombinationofmCherrywithavarietyofDnaAfragments,wedemonstrate that the membrane binding function is delocalized on the surface of the protein’s domain III, rather than confined to a particular sequence. We propose a new binding-bending mechanism to explain the membrane-induced nucleotide releasefromDnaA.Thismechanismwouldbefundamentaltotheinitiationofreplication. Citation:RegevT,MyersN,ZarivachR,FishovI(2012)AssociationoftheChromosomeReplicationInitiatorDnaAwiththeEscherichiacoliInnerMembraneIn Vivo:QuantityandModeofBinding.PLoSONE7(5):e36441.doi:10.1371/journal.pone.0036441 Editor:DipankarChatterji,IndianInstituteofScience,India ReceivedJanuary8,2012;AcceptedApril2,2012;PublishedMay4,2012 Copyright: ? 2012 Regev et al. This is an o