Reversal of PCNA Ubiquitylation by Ubp10 in Saccharomyces cerevisiae 英文参考文献.docVIP

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Reversal of PCNA Ubiquitylation by Ubp10 in Saccharomyces cerevisiae 英文参考文献.doc

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Reversal of PCNA Ubiquitylation by Ubp10 in Saccharomyces cerevisiae 英文参考文献

ReversalofPCNAUbiquitylationbyUbp10in Saccharomycescerevisiae AlfonsoGallego-Sa′nchez1,SoniaAndre′s1,FranciscoConde2,PedroA.San-Segundo2,AvelinoBueno1,3* 1InstitutodeBiolog?′aMolecularyCelulardelCa′ncer, UniversidaddeSalamanca/CSIC,Salamanca,Spain,2InstitutodeBiolog?′aFuncionalyGeno′mica,Universidadde Salamanca/CSIC,Salamanca,Spain,3DepartamentodeMicrobiolog?′ayGene′tica,UniversidaddeSalamanca/CSIC,Salamanca,Spain Abstract Regulation of PCNA ubiquitylation plays a key role in the tolerance to DNA damage in eukaryotes. Although the evolutionary conserved mechanism of PCNA ubiquitylation is well understood, the deubiquitylation of ubPCNA remains poorly characterized. Here, we show that thehistone H2BK123 ubiquitin protease Ubp10 also deubiquitylates ubPCNAin Saccharomycescerevisiae.OurresultssustainthatUbp10-dependentdeubiquitylationoftheslidingclampPCNAnormally takesplace during S phase,likely inresponse tothe simplepresence of ubPCNA. In agreementwith this, weshow that Ubp10formsacomplexwithPCNAinvivo.Interestingly,wealsoshowthatdeletionofUBP10altersindifferentwaysthe interactionofPCNAwithDNApolymerasef–associatedproteinRev1andwithaccessorysubunitRev7.Whiledeletionof UBP10enhancesPCNA–Rev1interaction,itdecreasessignificantlyRev7bindingtotheslidingclamp.Finally,wereportthat Ubp10counteractsRad18E3-ubiquitinligaseactivityonPCNAatlysine164insuchamannerthatderegulationofUbp10 expressioncausestoleranceimpairmentandMMShypersensitivity. Citation:Gallego-Sa′nchez A,Andre′s S,CondeF,San-SegundoPA,BuenoA(2012)ReversalofPCNAUbiquitylationbyUbp10inSaccharomycescerevisiae .PLoS Genet8(7):e1002826.doi:10.1371/journal.pgen.1002826 Editor:NancyMaizels,UniversityofWashington,UnitedStatesofAmerica ReceivedNovember21,2011;AcceptedMay25,2012;PublishedJuly19,2012 Copyright:?2012Gallego-Sa′nchezetal.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorands