Replication Fork Reversal after Replication–Transcription Collision 英文参考文献.docVIP

ReplicationForkReversalafterReplication–Transcription Collision AnneL.DeSeptenville1,2,Ste′phaneDuigou1,2.,HasnaBoubakri1,2.¤,Be′ne′dicteMichel1,2 * 1CNRS,CentredeGe′ne′tiqueMole′culaire,UPR3404,Gif-sur-Yvette,France,2Universite′ Paris-Sud,Orsay,France Abstract Replicationforkarrestisarecognizedsourceofgeneticinstability,andtranscriptionisoneofthemostprominentcausesof replicationimpediment.WeanalyzeheretherequirementforrecombinationproteinsinEscherichiacoliwhenreplication– transcriptionhead-oncollisionsareinducedataspecificsitebytheinversionofahighlyexpressedribosomaloperon(rrn). RecBC is the only recombination protein required for cell viability under these conditions of increased replication- transcription collisions. In its absence, fork breakage occurs at the site of collision, and the resulting linear DNA is not repairedandisslowlydegradedbytheRecJexonuclease.LethalforkbreakageisalsoobservedincellsthatlackRecAand RecD, i.e. when both homologous recombination and the potent exonuclease V activity of the RecBCD complex are inactivated,withaslowdegradationoftheresultinglinearDNAbythecombinedactionoftheRecBChelicaseandtheRecJ exonuclease.ThesizesofthemajorlinearfragmentsindicatethatDNAdegradationissloweddownbytheencounterwith anotherrrnoperon.TheamountoflinearDNAdecreasesnearlytwo-foldwhentheHollidayjunctionresolvaseRuvABCis inactivated in recB, as well as in recA recD mutants, indicating that part of the linear DNA is formed by resolution of a Hollidayjunction.Ourresultssuggestthatreplicationforkreversaloccursafterreplication–transcriptionhead-oncollision, andweproposethatitpromotestheactionoftheaccessoryreplicativehelicasesthatdislodgetheobstacle. Citation:DeSeptenvilleAL,DuigouS,BoubakriH,MichelB(2012)ReplicationForkReversalafterReplication–TranscriptionCollision.PLoSGenet8(4):e1002622. doi:10.1371/journal.pgen.1002622 Editor:WilliamF.Burkholder,AgencyforScience,Technology,andResearch,Singapore ReceivedNovember15,2011;AcceptedFebruary10,2012;PublishedApri