title: The Arabidopsis thaliana mitogen-activated protein kinases MPK3 and MPK6 target a subclass of 'VQ-motif'-containing proteins to regulate immune responses.
abstract: Mitogen-activated protein kinase (MAPK) cascades play key roles in plant immune signalling, and elucidating their regulatory functions requires the identification of the pathway-specific substrates. We used yeast two-hybrid interaction screens, in vitro kinase assays and mass spectrometry-based phosphosite mapping to study a family of MAPK substrates. Site-directed mutagenesis and promoter-reporter fusion studies were performed to evaluate the impact of substrate phosphorylation on downstream signalling. A subset of the Arabidopsis thaliana VQ-motif-containing proteins (VQPs) were phosphorylated by the MAPKs MPK3 and MPK6, and renamed MPK3/6-targeted VQPs (MVQs). When plant protoplasts (expressing these MVQs) were treated with the flagellin-derived peptide flg22, several MVQs were destabilized in vivo. The MVQs interact with specific WRKY transcription factors. Detailed analysis of a representative member of the MVQ subset, MVQ1, indicated a negative role in WRKY-mediated defence gene expression - with mutation of the VQ-motif abrogating WRKY binding and causing mis-regulation of defence gene expression. We postulate the existence of a variety of WRKY-VQP-containing transcriptional regulatory protein complexes that depend on spatio-temporal VQP and WRKY expression patterns. Defence gene transcription can be modulated by changing the composition of these complexes - in part - through MAPK-mediated VQP degradation.
authors.affiliation: Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120, Halle, Germany.
authors.forename: Pascal
authors.initials: P
authors.lastname: Pecher
authors.affiliation: Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120, Halle, Germany.
authors.forename: Lennart
authors.initials: L
authors.lastname: Eschen-Lippold
authors.affiliation: Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120, Halle, Germany.
authors.forename: Siska
authors.initials: S
authors.lastname: Herklotz
authors.affiliation: Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120, Halle, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Kuhle
authors.affiliation: Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120, Halle, Germany.
authors.forename: Kai
authors.initials: K
authors.lastname: Naumann
authors.affiliation: Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120, Halle, Germany.
authors.forename: Gerit
authors.initials: G
authors.lastname: Bethke
authors.affiliation: Department of Plant Molecular Biology and Physiology, Georg August University of Goettingen, Julia-Lermontowa-Weg 3, D-37077, Goettingen, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Uhrig
authors.affiliation: Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120, Halle, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Weyhe
authors.affiliation: Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120, Halle, Germany.
authors.forename: Dierk
authors.initials: D
authors.lastname: Scheel
authors.affiliation: Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120, Halle, Germany.
authors.forename: Justin
authors.initials: J
authors.lastname: Lee
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000276:immunology* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D005408:Flagellin / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D020929:Mitogen-Activated Protein Kinase Kinases / Q000235:genetics / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D010935:Plant Diseases / Q000276:immunology; D030821:Plants, Genetically Modified; D055550:Protein Stability; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; C486389:NHL10 protein, Arabidopsis; D014157:Transcription Factors; D005408:Flagellin; C458516:AtMPK3 protein, Arabidopsis; C436293:MPK6 protein, Arabidopsis; D020928:Mitogen-Activated Protein Kinases; D020929:Mitogen-Activated Protein Kinase Kinases
keywords: VQ-motif; defence; immunity; mitogen-activated protein kinase (MAPK); pathogen-associated molecular pattern (PAMP); phosphorylation; signalling
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