Rhodococcus equi’s Extreme Resistance to Hydrogen Peroxide Is Mainly Conferred by One of Its Four Catalase Genes 英文参考文献.docVIP

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Rhodococcus equi’s Extreme Resistance to Hydrogen Peroxide Is Mainly Conferred by One of Its Four Catalase Genes 英文参考文献.doc

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Rhodococcus equi’s Extreme Resistance to Hydrogen Peroxide Is Mainly Conferred by One of Its Four Catalase Genes 英文参考文献

Rhodococcusequi’sExtremeResistancetoHydrogen PeroxideIsMainlyConferredbyOneofItsFourCatalase Genes PaulineBidaud1.,LaurentHe′bert1.,CorinneBarbey1¤a,Anne-Ce′cileAppourchaux1¤b,RiccardoTorelli2, MaurizioSanguinetti2,ClaireLaugier1,SandrinePetry1* 1Dozule′LaboratoryforEquineDiseases,UnitBacteriologyandParasitology,ANSES,Goustranville,France,2InstituteofMicrobiology,CatholicUniversityofSacredHeart, Rome,Italy Abstract Rhodococcus equi is one of the most widespread causes of disease in foals aged from 1 to 6months. R. equi possesses antioxidant defense mechanisms to protect it from reactive oxygen metabolites such as hydrogen peroxide (H2O2) generatedduringtherespiratoryburstofphagocyticcells.Thesedefensemechanismsincludeenzymessuchascatalase, whichdetoxifyhydrogenperoxide.Recently,ananalysisoftheR.equi103genomesequencerevealedthepresenceoffour potential catalase genes. We first constructed DkatA-, DkatB-, DkatC-and DkatD-deficient mutants to study the ability of R.equitosurviveexposuretoH2O2invitroandwithinmouseperitonealmacrophages.ResultsshowedthatDkatAand,toa lesserextentDkatC,wereaffectedby80mMH2O2.Moreover,katAdeletionseemstosignificantlyaffecttheabilityofR.equi to survive within murine macrophages. We finally investigated the expression of the four catalases in response to H2O2 assays with a real time PCR technique. Results showed that katA is overexpressed 367.9times (6122.6) in response to exposureto50mMofH2O2addedinthestationaryphase,and3.11times(60.59)whentreatmentwasadministeredinthe exponentialphase.Inuntreatedbacteria,katB,katCandkatDwereoverexpressedfrom4.3to17.5timesinthestationary compared to the exponential phase. Taken together, our results show that KatA is the major catalase involved in the extremeH2O2resistancecapabilityofR.equi. Citation:BidaudP,He′bertL,BarbeyC,AppourchauxA-C,TorelliR,etal.(2012)Rhodococcusequi’sExtremeResistancetoHydrogenPeroxideIsMainlyConferred byOneofItsFourCatalaseGenes.PLoSONE7(8):e42396.doi:10.1371/journal.pone.0042396