Screening for Toxic Amyloid in Yeast Exemplifies the Role of Alternative Pathway Responsible for Cytotoxicity 英文参考文献.docVIP

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Screening for Toxic Amyloid in Yeast Exemplifies the Role of Alternative Pathway Responsible for Cytotoxicity 英文参考文献.doc

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Screening for Toxic Amyloid in Yeast Exemplifies the Role of Alternative Pathway Responsible for Cytotoxicity 英文参考文献

ScreeningforToxicAmyloidinYeastExemplifiesthe RoleofAlternativePathwayResponsibleforCytotoxicity JulienCouthouis1.,KarineRe′bora1.,Franc?oiseImmel1,KarineBerthelot1,MichelCastroviejo2, ChristopheCullin1* 1IBGC,UMR5095,CNRSUniversite′ Bordeaux2‘‘VictorSegalen’’,Bordeaux,France,2REGER,UMR5097CNRSUniversite′ Bordeaux2‘‘VictorSegalen’’,Bordeaux,France Abstract The relationship between amyloid and toxic species is a central problem since the discovery of amyloid structures in differentdiseases.Despiteintensiveeffortsinthefield,thedeleteriousspeciesremainsunknownatthemolecularlevel.This may reflect thelack ofanystructure-toxicity study based on ageneticapproach. Hereweshowthatastructure-toxicity study without any biochemical prerequisite can be successfully achieved in yeast. A PCR mutagenesis of the amyloid domainofHET-sleadstotheidentificationofamutantthatmightimpaircellularviability.Cellularandbiochemicalanalyses demonstratethatthistoxicmutantformsGFP-amyloidaggregatesthatdifferfromthewild-typeaggregatesintheirshape, sizeandmolecularorganization.ThechaperoneHsp104thathelpstodisassembleproteinaggregatesisstrictlyrequiredfor thecellulartoxicity.Ourstructure-toxicitystudysuggeststhatthesmallestaggregatesarethemosttoxic,andopensanew waytoanalyzetherelationshipbetweenstructureandtoxicityofamyloidspecies. Citation:CouthouisJ,Re′boraK,ImmelF,BerthelotK,CastroviejoM,etal.(2009)ScreeningforToxicAmyloidinYeastExemplifiestheRoleofAlternativePathway ResponsibleforCytotoxicity.PLoSONE4(3):e4539.doi:10.1371/journal.pone.0004539 Editor:RichardMayeux,ColumbiaUniversity,UnitedStatesofAmerica ReceivedOctober17,2008;AcceptedFebruary2,2009;PublishedMarch5,2009 Copyright: ? 2009 Couthouis et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:TheworkwassupportedbygrantsfromGIS‘‘Maladierares’’andANRProje