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

Mol Plant. 2008 May;1(3):401-10. doi: 10.1093/mp/ssn009. Epub 2008 Mar 21.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptotic Protease-Activating Factor 1 / chemistry
  • Genetic Complementation Test*
  • Humans
  • Hypersensitivity / genetics
  • Hypersensitivity / immunology
  • Lycopersicon esculentum / genetics*
  • Lycopersicon esculentum / immunology
  • Lycopersicon esculentum / metabolism
  • Models, Molecular
  • Mutation
  • Phenotype
  • Plant Diseases / genetics
  • Plant Diseases / immunology
  • Plant Leaves / genetics
  • Plant Proteins / chemistry
  • Plant Proteins / genetics*
  • Plant Proteins / immunology
  • Point Mutation
  • Promoter Regions, Genetic
  • Protein Conformation
  • Signal Transduction

Substances

  • Apoptotic Protease-Activating Factor 1
  • Plant Proteins