title: Direct regulation of the NADPH oxidase RBOHD by the PRR-associated kinase BIK1 during plant immunity.
abstract: The rapid production of reactive oxygen species (ROS) burst is a conserved signaling output in immunity across kingdoms. In plants, perception of pathogen-associated molecular patterns (PAMPs) by surface-localized pattern recognition receptors (PRRs) activates the NADPH oxidase RBOHD by hitherto unknown mechanisms. Here, we show that RBOHD exists in complex with the receptor kinases EFR and FLS2, which are the PRRs for bacterial EF-Tu and flagellin, respectively. The plasma-membrane-associated kinase BIK1, which is a direct substrate of the PRR complex, directly interacts with and phosphorylates RBOHD upon PAMP perception. BIK1 phosphorylates different residues than calcium-dependent protein kinases, and both PAMP-induced BIK1 activation and BIK1-mediated phosphorylation of RBOHD are calcium independent. Importantly, phosphorylation of these residues is critical for the PAMP-induced ROS burst and antibacterial immunity. Our study reveals a rapid regulatory mechanism of a plant RBOH, which occurs in parallel with and is essential for its paradigmatic calcium-based regulation.
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK; RIKEN Center for Sustainable Resource Science, Plant Immunity Research Group, Suehiro-cho 1-7-22 Tsurumi-ku, Yokohama 230-0045, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kadota
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK.
authors.forename: Jan
authors.initials: J
authors.lastname: Sklenar
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK.
authors.forename: Paul
authors.initials: P
authors.lastname: Derbyshire
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK.
authors.forename: Lena
authors.initials: L
authors.lastname: Stransfeld
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK; RIKEN Center for Sustainable Resource Science, Plant Immunity Research Group, Suehiro-cho 1-7-22 Tsurumi-ku, Yokohama 230-0045, Japan.
authors.forename: Shuta
authors.initials: S
authors.lastname: Asai
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK.
authors.forename: Vardis
authors.initials: V
authors.lastname: Ntoukakis
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK.
authors.forename: Jonathan Dg
authors.initials: JD
authors.lastname: Jones
authors.affiliation: RIKEN Center for Sustainable Resource Science, Plant Immunity Research Group, Suehiro-cho 1-7-22 Tsurumi-ku, Yokohama 230-0045, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Shirasu
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK.
authors.forename: Frank
authors.initials: F
authors.lastname: Menke
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Jones
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK. Electronic address: cyril.zipfel@tsl.ac.uk.
authors.forename: Cyril
authors.initials: C
authors.lastname: Zipfel
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics / Q000276:immunology / Q000382:microbiology; D029681:Arabidopsis Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D002460:Cell Line; D004789:Enzyme Activation; D005408:Flagellin / Q000276:immunology / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D007113:Immunity, Innate; D008024:Ligands; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D019255:NADPH Oxidases / Q000235:genetics / Q000276:immunology*; D010444:Peptide Elongation Factor Tu / Q000276:immunology / Q000378:metabolism; D010766:Phosphorylation; D057865:Plant Immunity; D054046:Plant Stomata / Q000276:immunology / Q000378:metabolism; D011494:Protein Kinases / Q000276:immunology / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D011971:Receptors, Immunologic / Q000378:metabolism; D051192:Receptors, Pattern Recognition / Q000276:immunology / Q000378:metabolism; D015398:Signal Transduction; D014026:Nicotiana / Q000201:enzymology* / Q000235:genetics / Q000276:immunology / Q000382:microbiology
chemical_list: D029681:Arabidopsis Proteins; C545106:EFR protein, Arabidopsis; D008024:Ligands; D009097:Multienzyme Complexes; D017382:Reactive Oxygen Species; D011971:Receptors, Immunologic; D051192:Receptors, Pattern Recognition; D005408:Flagellin; C568857:AT5G47910 protein, Arabidopsis; D019255:NADPH Oxidases; D011494:Protein Kinases; C411853:FLS2 protein, Arabidopsis; C509522:BIK1 protein, Arabidopsis; D017346:Protein Serine-Threonine Kinases; D010444:Peptide Elongation Factor Tu