Disruption of Yarrowia lipolytica TPS1 Gene Encoding Trehalose-6-P Synthase Does Not Affect Growth in Glucose but Impairs Growth at High Temperature 英文参考文献.docVIP

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Disruption of Yarrowia lipolytica TPS1 Gene Encoding Trehalose-6-P Synthase Does Not Affect Growth in Glucose but Impairs Growth at High Temperature 英文参考文献.doc

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Disruption of Yarrowia lipolytica TPS1 Gene Encoding Trehalose-6-P Synthase Does Not Affect Growth in Glucose but Impairs Growth at High Temperature 英文参考文献

DisruptionofYarrowialipolyticaTPS1GeneEncoding Trehalose-6-PSynthaseDoesNotAffectGrowthin GlucosebutImpairsGrowthatHighTemperature Carmen-LissetFlores1*,CarlosGancedo1,ThomasPetit2 1DepartmentofMetabolismandCellSignalling,InstitutodeInvestigacionesBiome′dicas‘‘AlbertoSols’’CSIC-UAM,Madrid,Spain,2Universite′deLaRe′union,Laboratoire deChimiedesSubstancesNaturellesetdesSciencesdesAliments,InstitutUniversitairedeTechnologie,De′partementGe′nie Biologique,SaintPierre,France Abstract We have cloned the Yarrowia lipolytica TPS1 gene encoding trehalose-6-P synthase by complementation of the lack of growthinglucoseofaSaccharomycescerevisiaetps1mutant.DisruptionofYlTPS1couldonlybeachievedwithacassette placedinthe39halfofitscodingregionduetotheoverlapofitssequencewiththepromoteroftheessentialgeneYlTFC1. TheYltps1mutantgrewinglucosealthoughtheY.lipolyticahexokinaseisextremelysensitivetoinhibitionbytrehalose-6-P. The presence of a glucokinase, insensitive to trehalose-6-P, that constitutes about 80% of the glucose phosphorylating capacityduringgrowthinglucosemayaccountforthegrowthphenotype.Trehalosecontentwasbelow1nmol/mgdry weight in Y. lipolytica, but it increased in strains expressing YlTPS1 under the control of the YlTEF1promoter or with a disruptionofYALI0D15598encodingaputativetrehalase.mRNAlevelsofYlTPS1werelowanddidnotrespondtothermal stresses,butthatofYlTPS2(YALI0D14476)andYlTPS3(YALI0E31086)increased4and6times,repectively,byheattreatment. Disruption of YlTPS1 drastically slowed growth at 35uC. Homozygous Yltps1 diploids showed a decreased sporulation frequencythatwasascribedtothelowlevelofYALI0D20966mRNAanhomologoftheS.cerevisiaeMCK1whichencodesa proteinkinasethatactivatesearlymeioticgeneexpression. Citation:FloresC-L,GancedoC,PetitT(2011)DisruptionofYarrowialipolyticaTPS1GeneEncodingTrehalose-6-PSynthaseDoesNotAffectGrowthinGlucose butImpairsGrowthatHighTemperature.PLoSONE6(9):e23695.doi:10.1371/journal.pone.0023695 Editor:RobertAlanArkowitz,InstituteofDevelo