Simple Sequence Repeats and Mucoid Conversion Biased mucA Mutagenesis in Mismatch Repair-Deficient Pseudomonas aeruginosa 英文参考文献.docVIP

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Simple Sequence Repeats and Mucoid Conversion Biased mucA Mutagenesis in Mismatch Repair-Deficient Pseudomonas aeruginosa 英文参考文献.doc

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Simple Sequence Repeats and Mucoid Conversion Biased mucA Mutagenesis in Mismatch Repair-Deficient Pseudomonas aeruginosa 英文参考文献

SimpleSequenceRepeatsandMucoidConversion: BiasedmucAMutagenesisinMismatchRepair-Deficient Pseudomonasaeruginosa AlejandroJ.Moyano,AndreaM.Smania* Centro de Investigaciones en Qu?′mica Biolo′gica de Co′rdoba (CIQUIBIC), CONICET, Departamento de Qu?′mica Biolo′gica, Facultad de Ciencias Qu?′micas, Universidad NacionaldeCo′rdoba,Co′rdoba,Argentina Abstract InPseudomonasaeruginosa,conversiontothemucoidphenotypemarkstheonsetofanirreversiblestateoftheinfectionin CysticFibrosis(CF)patients.ThemainpathwayformucoidconversionismutagenesisofthemucAgene,frequentlydueto 21bpdeletionsinasimplesequencerepeat(SSR)of5Gs(G5-SSR426).WehaverecentlyobservedthatthismucAmutation is particularly accentuated in Mismatch Repair System (MRS)-deficient cells grown in vitro. Interestingly, previous reports haveshownahighprevalenceofhypermutableMRS-deficientstrainsoccurringnaturallyinCFchroniclunginfections.Here, weusedmucAasaforwardmutationmodeltosystematicallyevaluatetheroleofG5-SSR426inconversiontomucoidyina MRS-deficientbackground,withthisbeingthefirstanalysiscombiningSSR-dependentlocalizedhypermutabilityandthe acquisition of a particular virulence/persistence trait in P. aeruginosa. In this study, mucA alleles were engineered with differentcontentsofG:CSSRs,andtestedfortheireffectonthemucoidconversionfrequencyandmucAmutationalspectra inamutS-deficientstrainofP.aeruginosa.Importantly,deletionofG5-SSR426severelyreducedtheemergencefrequencyof mucoidvariants,withnopreferentialsiteofmutagenesiswithinmucA.Moreover,althoughmutagenesisinmucAwasnot totallyremoved,thiswasnolongerthemainpathwayformucoidconversion,suggestingthatG5-SSR426biasedmutations towards mucA. Mutagenesis in mucA was restored by the addition of a new SSR (C6-SSR431), and even synergistically increasedwhenG5-SSR426andC6-SSR431werepresentsimultaneously,withthemucAmutationsbeingrestrictedto2 1bp deletionswithinanyofbothG:CSSRs.TheseresultsconfirmacriticalroleforG5-SSR426enhancingthemutagenicprocessof mucA in MRS-deficient cells, a