title: Transcomplementation, but not physical association of the CC-NB-ARC and LRR domains of tomato R protein Mi-1.2 is altered by mutations in the ARC2 subdomain.
abstract: Race-specific disease resistance in plants is mediated by Resistance (R) proteins that recognize pathogen attack and initiate defence responses. Most R proteins contain a central NB-ARC domain and a C-terminal leucine-rich repeat (LRR) domain. We analyzed the intramolecular interaction of the LRR domain of tomato R protein Mi-1.2 with its N-terminus. We expressed the CC-NB-ARC and LRR parts in trans and analyzed functional transcomplementation and physical interactions. We show that these domains functionally transcomplement when expressed in trans. Known autoactivating LRR domain swaps were found to induce a hypersensitive response (HR) upon co-expression. Likewise, autoactivating mutants in the NB subdomain transcomplemented to induce HR. Point mutations in the ARC2 subdomain that induce strong autoactivation in the full-length Mi-1.2 protein, however, fail to induce HR in the transcomplementation assay. These data indicate distinct functions for the NB-ARC subdomains in induction of HR signalling. Furthermore, dissociation of the LRR is not required to release its negative regulation, as in all combinations of CC-NB-ARC and LRR domains tested, a physical interaction was observed.
authors.affiliation: Plant Pathology, Swammerdam Institute for Life Sciences, University of Amsterdam, PO Box 94062, 1090 GB Amsterdam, the Netherlands.
authors.forename: Gerben
authors.initials: G
authors.lastname: van Ooijen
authors.forename: Gabriele
authors.initials: G
authors.lastname: Mayr
authors.forename: Mario
authors.initials: M
authors.lastname: Albrecht
authors.forename: Ben J C
authors.initials: BJ
authors.lastname: Cornelissen
authors.forename: Frank L W
authors.initials: FL
authors.lastname: Takken
mesh_terms: D053477:Apoptotic Protease-Activating Factor 1 / Q000737:chemistry; D005816:Genetic Complementation Test; D006801:Humans; D006967:Hypersensitivity / Q000235:genetics / Q000276:immunology; D018551:Solanum lycopersicum / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D008958:Models, Molecular; D009154:Mutation; D010641:Phenotype; D010935:Plant Diseases / Q000235:genetics / Q000276:immunology; D018515:Plant Leaves / Q000235:genetics; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D017354:Point Mutation; D011401:Promoter Regions, Genetic; D011487:Protein Conformation; D015398:Signal Transduction
chemical_list: D053477:Apoptotic Protease-Activating Factor 1; D010940:Plant Proteins