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1、InteractionsbetweentheS-DomainReceptorKinasesandAtPUB-ARME3UbiquitinLigasesSuggestaConservedSignalingPathwayinArabidopsis1[W][OA]22,3MarcusA.Samuel,YashwantiMudgil,JenniferN.Salt,Fre´de´ricDelmas,ShalinyRamachandran,AndreaChilelli,andDaphneR.Goring*DepartmentofCella
2、ndSystemsBiology,UniversityofToronto,Toronto,Ontario,CanadaM5S3B2TheArabidopsis(Arabidopsisthaliana)genomeencompassesmultiplereceptorkinasefamilieswithhighlyvariableextracellulardomains.Despitetheirlargenumbers,thevariousligandsandthedownstreaminteractingpartnersfor
3、thesekinaseshavebeendecipheredonlyforafewmembers.Onesuchmember,theS-receptorkinase,isknowntomediatetheself-incompatibility(SI)responseinBrassica.S-receptorkinasehasbeenshowntointeractandphosphorylateaU-box/ARM-repeat-containingE3ligase,ARC1,which,inturn,actsasaposit
4、iveregulatoroftheSIresponse.InanefforttoidentifyconservedsignalingpathwaysinArabidopsis,weperformedyeasttwo-hybridanalysesofvariousS-domainreceptorkinasefamilymemberswithrepresentativeArabidopsisplantU-box/ARM-repeat(AtPUB-ARM)E3ligases.ThekinasedomainsfromS-domainr
5、eceptorkinaseswerefoundtointeractwithARM-repeatdomainsfromAtPUB-ARMproteins.Thesekinasedomains,alongwithM-locusproteinkinase,apositiveregulatorofSIresponse,werealsoabletophosphorylatetheARM-repeatdomainsininvitrophosphorylationassays.Subcellularlocalizationpatternsw
6、ereinvestigatedusingtransientexpressionassaysintobacco(Nicotianatabacum)BY-2cellsandchangesweredetectedinthepresenceofinteractingkinases.Finally,potentiallinkstotheinvolvementoftheseinteractingmodulestothehormoneabscisicacid(ABA)wereinvestigated.Interestingly,AtPUB9
7、displayedredistributiontotheplasmamembraneofBY-2cellswheneithertreatedwithABAorcoexpressedwiththeactivekinasedomainofARK1.Aswell,T-DNAinsertionmutantsforARK1andAtPUB9lineswerealteredintheirABAsensitivityduringgerminationandactedatorupstreamofABI3,indicatingpotential
8、involvementoftheseproteinsinABAresponses.Theprocessofubiquitin-mediatedproteindegrada-theArabidopsis(Arabidopsisthaliana)genome(Krafttioni
