CRK2 and C-terminal phosphorylation of NADPH oxidase RBOHD regulate ROS production in Arabidopsis
CRK2 and C-terminal phosphorylation of NADPH oxidase RBOHD regulate ROS production in Arabidopsis
ABSTRACT Reactive oxygen species (ROS) are important messengers in eukaryotic organisms and their production is tightly controlled. Active extracellular ROS production by NADPH oxidases in plants is triggered by receptor-like protein kinase (RLK)-dependent signaling networks. Here we show that the cysteine-rich RLK CRK2 kinase activity is required for plant growth and CRK2 exists in a preformed complex with the NADPH oxidase RBOHD in Arabidopsis. Functional CRK2 is required for the full elicitor-induced ROS burst and consequently the crk2 mutant is impaired in defense against the bacterial pathogen Pseudomonas syringae pv. tomato DC3000. Our work demonstrates that CRK2 regulates plant innate immunity. We identified in vitro CRK2-dependent phosphorylation sites in the C-terminal region of RBOHD. Phosphorylation of S703 RBOHD is enhanced upon flg22 treatment and substitution of S703 with alanine reduced ROS production in Arabidopsis. Phylogenetic analysis suggests that phospho-sites in C-terminal region of RBOHD are conserved throughout the plant lineage and between animals and plants. We propose that regulation of NADPH oxidase activity by phosphorylation of the C-terminal region might be an ancient mechanism and that CRK2 is an important element in regulating MAMP-triggered ROS production. One-sentence summaryCRK2 associates with and activates RBOHD to trigger MAMP-induced ROS production and reveals a novel regulatory mechanism for plant NADPH oxidases through phosphorylation of the C-terminus.
Nakagami Hirofumi、Wrzaczek Michael、Harzen Anne、Kimura Sachie、Hunter Kerri、Tran Huy Cuong、Vaattovaara Aleksia、Vaahtera Lauri、Mei?ner Lena、Toyota Masatsugu、Rokka Anne、Wilkens Maya Melina Tabea、Stolze Sara Christina、Hamann Thorsten、Citterico Matteo
Protein Mass Spectrometry Group, Max-Planck Institute for Plant Breeding ResearchOrganismal and Evolutionary Biology Research Programme, Viikki Plant Science Centre, Faculty of Biological and Environmental Sciences, University of HelsinkiProtein Mass Spectrometry Group, Max-Planck Institute for Plant Breeding ResearchOrganismal and Evolutionary Biology Research Programme, Viikki Plant Science Centre, Faculty of Biological and Environmental Sciences, University of Helsinki||Ritsumeikan Global Innovation Research Organization, Ritsumeikan UniversityOrganismal and Evolutionary Biology Research Programme, Viikki Plant Science Centre, Faculty of Biological and Environmental Sciences, University of HelsinkiOrganismal and Evolutionary Biology Research Programme, Viikki Plant Science Centre, Faculty of Biological and Environmental Sciences, University of Helsinki||Department of Biology, Lund UniversityOrganismal and Evolutionary Biology Research Programme, Viikki Plant Science Centre, Faculty of Biological and Environmental Sciences, University of HelsinkiDepartment of Biology, Norwegian University of Science and TechnologyOrganismal and Evolutionary Biology Research Programme, Viikki Plant Science Centre, Faculty of Biological and Environmental Sciences, University of Helsinki||Technische Universit?t BraunschweigDepartment of Biochemistry and Molecular Biology, Saitama University||Department of Botany, University of WisconsinTurku Centre for Biotechnology, University of Turku and ?bo AkademiOrganismal and Evolutionary Biology Research Programme, Viikki Plant Science Centre, Faculty of Biological and Environmental Sciences, University of Helsinki||Institute of Molecular Biology gGmbH (IMB)Protein Mass Spectrometry Group, Max-Planck Institute for Plant Breeding ResearchDepartment of Biology, Norwegian University of Science and TechnologyOrganismal and Evolutionary Biology Research Programme, Viikki Plant Science Centre, Faculty of Biological and Environmental Sciences, University of Helsinki
植物学分子生物学生物化学
Nakagami Hirofumi,Wrzaczek Michael,Harzen Anne,Kimura Sachie,Hunter Kerri,Tran Huy Cuong,Vaattovaara Aleksia,Vaahtera Lauri,Mei?ner Lena,Toyota Masatsugu,Rokka Anne,Wilkens Maya Melina Tabea,Stolze Sara Christina,Hamann Thorsten,Citterico Matteo.CRK2 and C-terminal phosphorylation of NADPH oxidase RBOHD regulate ROS production in Arabidopsis[EB/OL].(2025-03-28)[2025-04-28].https://www.biorxiv.org/content/10.1101/618819.点此复制
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