Sustained mitogen-activated protein kinase activation reprograms defense metabolism and phosphoprotein profile in Arabidopsis thaliana

Lassowskat, Ines; Böttcher, Christoph GND; Eschen-Lippold, Lennart; Scheel, Dierk; Lee, Justin

Mitogen-activatedproteinkinases(MAPKs)targetavarietyofproteinsubstratestoregulatecellularsignalingprocessesineukaryotes.Inplants,thenumberofidentifiedMAPKsubstratesthatcontrolplantdefenseresponsesisstilllimited.Here,wegeneratedtransgenicArabidopsisthalianaplantswithaninduciblesystemtosimulateinvivoactivationoftwostress-activatedMAPKs,MPK3,andMPK6.MetabolomeanalysisrevealedthatthisartificialMPK3/6activation(withoutanyexposuretopathogensorotherstresses)issufficienttodrivetheproductionofmajordefense-relatedmetabolites,includingvariouscamalexin,indoleglucosinolateandagmatinederivatives.Anaccompanying(phospho)proteomeanalysisledtodetectionofhundredsofpotentialphosphoproteinsdownstreamofMPK3/6activation.BesidesknownMAPKsubstrates,manycandidatesonthislistpossesstypicalMAPK-targetedphosphositesandinmanycases,thecorrespondingphosphopeptidesweredetectedbymassspectrometry.Notably,severaloftheseputativephosphoproteinshavebeenreportedtobeassociatedwiththebiosynthesisofantimicrobialdefensesubstances(e.g.,WRKYtranscriptionfactorsandproteinsencodedbythegenesfromthe“PEN”pathwayrequiredforpenetrationresistancetofilamentouspathogens).Thus,thisworkprovidesaninventoryofcandidatephosphoproteins,includingputativedirectMAPKsubstrates,forfutureanalysisofMAPK-mediateddefensecontrol.(ProteomicsdataareavailablewiththeidentifierPXD001252viaProteomeXchange,http://proteomecentral.proteomexchange.org).

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Lassowskat, Ines / Böttcher, Christoph / Eschen-Lippold, Lennart / et al: Sustained mitogen-activated protein kinase activation reprograms defense metabolism and phosphoprotein profile in Arabidopsis thaliana. 2014.

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