title: A patch of surface-exposed residues mediates negative regulation of immune signaling by tomato Pto kinase.
abstract: Tomato (Lycopersicon esculentum) Pto kinase specifically recognizes the Pseudomonas effector proteins AvrPto and AvrPtoB, leading to induction of defense responses and hypersensitive cell death. Structural modeling of Pto combined with site-directed mutagenesis identified a patch of surface-exposed residues required for native regulation of signaling. Mutations in this area resulted in constitutive gain-of-function (CGF) forms of Pto that activated AvrPto-independent cell death via the cognate signaling pathway. The patch overlaps the peptide binding region of the kinase catalytic cleft and is part of a broader region required for interaction with bacterial effectors. We propose that the negative regulatory patch is normally occupied by a peptide that represses Pto signaling. Furthermore, we found that Pto kinase activity was required for Avr-dependent activation but dispensable for signaling by CGF forms of Pto. This suggests that Pto signals by a conformational change rather than phosphorylation of downstream substrates in the defense signaling pathway.
authors.affiliation: Sainsbury Laboratory, Norwich Research Park, Norwich, Norfolk, NR4 7UH, United Kingdom.
authors.forename: Ai-Jiuan
authors.initials: AJ
authors.lastname: Wu
authors.forename: Vasilios M E
authors.initials: VM
authors.lastname: Andriotis
authors.forename: Marcus C
authors.initials: MC
authors.lastname: Durrant
authors.forename: John P
authors.initials: JP
authors.lastname: Rathjen
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D002384:Catalysis; D018551:Solanum lycopersicum / Q000201:enzymology* / Q000276:immunology; D009154:Mutation; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D010940:Plant Proteins; C084354:Pto protein, Lycopersicon; D017346:Protein Serine-Threonine Kinases