Evolution of Multidrug Resistance during Staphylococcus aureus Infection Involves Mutation of the Essential Two Component Regulator WalKR 英文参考文献.docVIP

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Evolution of Multidrug Resistance during Staphylococcus aureus Infection Involves Mutation of the Essential Two Component Regulator WalKR 英文参考文献.doc

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Evolution of Multidrug Resistance during Staphylococcus aureus Infection Involves Mutation of the Essential Two Component Regulator WalKR 英文参考文献

EvolutionofMultidrugResistanceduringStaphylococcus aureusInfectionInvolvesMutationoftheEssentialTwo ComponentRegulatorWalKR BenjaminP.Howden1,2,3,4*,ChristopherR.E.McEvoy1,DavidL.Allen4,KyraChua1,4,WeiGao2,4,PaulF. Harrison5,JanBell6,GeoffreyCoombs7,VickiBennett-Wood1,JessicaL.Porter1,RoyRobins-Browne1, JohnK.Davies4,TorstenSeemann5,TimothyP.Stinear1,4 1Department of Microbiology and Immunology, University of Melbourne, Victoria, Australia, 2Austin Centre for Infection Research (ACIR), Infectious Diseases Department, Austin Health, Heidelberg, Victoria, Australia, 3Microbiology Department, Austin Health, Heidelberg, Victoria, Australia, 4Department of Microbiology, Monash University, Clayton, Victoria, Australia, 5Victorian Bioinformatics Consortium, Monash University, Clayton, Victoria, Australia, 6SA Pathology, Womens and Children’sHospital,Adelaide,SouthAustralia,Australia,7MicrobiologyDepartment,RoyalPerthHospital,Perth,WesternAustralia,Australia Abstract AntimicrobialresistanceinStaphylococcusaureusisamajorpublichealththreat,compoundedbyemergenceofstrainswith resistance tovancomycin and daptomycin, both last lineantimicrobials. Here wehave performed high throughput DNA sequencing and comparative genomics for five clinical pairs of vancomycin-susceptible (VSSA) and vancomycin- intermediateST239S.aureus(VISA);eachpairisolatedbeforeandaftervancomycintreatmentfailure.Thesecomparisons revealedafrequentpatternofmutationamongtheVISAstrainswithintheessentialwalKRtwo-componentregulatorylocus involvedincontrolofcellwallmetabolism.Wethenconductedbi-directionalallelicexchangeexperimentsinourclinical VSSAandVISAstrainsandshowedthatsinglenucleotidesubstitutionswithineitherwalKorwalRleadtoco-resistanceto vancomycinanddaptomycin,andcausedthetypicalcellwallthickeningobservedinresistantclinicalisolates.IonTorrent genomesequencingconfirmednoadditionalregulatorymutationshadbeenintroducedintoeitherthewalRorwalKVISA mutants during the allelic exchange process. However, two potent