Molecular Characterization of Direct Target Genes and cis-Acting Consensus Recognized by Quorum-Sensing Regulator AphA in Vibrio parahaemolyticus 英文参考文献.docVIP

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Molecular Characterization of Direct Target Genes and cis-Acting Consensus Recognized by Quorum-Sensing Regulator AphA in Vibrio parahaemolyticus 英文参考文献.doc

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Molecular Characterization of Direct Target Genes and cis-Acting Consensus Recognized by Quorum-Sensing Regulator AphA in Vibrio parahaemolyticus 英文参考文献

MolecularCharacterizationofDirectTargetGenesand cis-ActingConsensusRecognizedbyQuorum-Sensing RegulatorAphAinVibrioparahaemolyticus FengjunSun1,2,YiquanZhang2,LiWang2,3,XiaojuanYan2,3,YafangTan2,ZhaobiaoGuo2,JingfuQiu3, RuifuYang2,PeiyuanXia1*,DongshengZhou2* 1Department of Pharmacy, Southwest Hospital, Third Military Medical University, Chongqing, People’s Republic of China, 2State Key Laboratory of Pathogen and Biosecurity,BeijingInstituteofMicrobiologyandEpidemiology,Beijing,People’sRepublicofChina,3SchoolofPublicHealth,ChongqingMedicalUniversity,Chongqing, People’sRepublicofChina Abstract Background: AphA is the master quorum-sensing (QS) regulator operating at low cell density in vibrios. Molecular regulationoftargetgenesbyAphAhasbeencharacterizedinVibrioharveyiandV.cholerae,butitisstillpoorlyunderstood inV.parahaemolyticus. Methodology/PrincipalFindings:TheAphAproteinsareextremelyconservedinV.parahaemolyticus,Vibriosp.Ex25,Vibrio sp.EJY3,V.harveyi,V.vulnificus,V.splendidus,V.anguillarum,V.cholerae,andV.furnissii.TheabovenineAphAorthologs appeartorecognizeconservedcis-actingDNAsignalswhichcanberepresentedbytwoconsensusconstructs,a20bpbox sequenceandapositionfrequencymatrix.V.parahaemolyticusAphArepressesthetranscriptionofahpA,qrr4,andopaR through direct AphA-target promoter DNA association, while it inhibits the qrr2-3 transcription in an indirect manner. Translation and transcription starts, core promoter elements for sigma factor recognition, Shine-Dalgarno sequences for ribosomerecognition,andAphA-bindingsites(containingcorrespondingAphAbox-likesequences)weredeterminedfor thethreedirectAphAtargetsahpA,qrr4,andopaRinV.parahaemolyticus. Conclusions/Significance:AphA-mediatedrepressionofahpA,qrr2-4,andopaRwascharacterizedinV.parahaemolyticusby using multiple biochemical and molecular experiments. The computational promoter analysis indicated the conserved mechanismoftranscriptionalregulationofQSregulator-encodinggenesahpA,qrr4,andopaRinvibrios. Citation:SunF,