Intramolecular interaction influences binding of the Flax L5 and L6 resistance proteins to their AvrL567 ligands

PLoS Pathog. 2012;8(11):e1003004. doi: 10.1371/journal.ppat.1003004. Epub 2012 Nov 29.

Abstract

L locus resistance (R) proteins are nucleotide binding (NB-ARC) leucine-rich repeat (LRR) proteins from flax (Linum usitatissimum) that provide race-specific resistance to the causal agent of flax rust disease, Melampsora lini. L5 and L6 are two alleles of the L locus that directly recognize variants of the fungal effector AvrL567. In this study, we have investigated the molecular details of this recognition by site-directed mutagenesis of AvrL567 and construction of chimeric L proteins. Single, double and triple mutations of polymorphic residues in a variety of AvrL567 variants showed additive effects on recognition strength, suggesting that multiple contact points are involved in recognition. Domain-swap experiments between L5 and L6 show that specificity differences are determined by their corresponding LRR regions. Most positively selected amino acid sites occur in the N- and C-terminal LRR units, and polymorphisms in the first seven and last four LRR units contribute to recognition specificity of L5 and L6 respectively. This further confirms that multiple, additive contact points occur between AvrL567 variants and either L5 or L6. However, we also observed that recognition of AvrL567 is affected by co-operative polymorphisms between both adjacent and distant domains of the R protein, including the TIR, ARC and LRR domains, implying that these residues are involved in intramolecular interactions to optimize detection of the pathogen and defense signal activation. We suggest a model where Avr ligand interaction directly competes with intramolecular interactions to cause activation of the R protein.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Basidiomycota / genetics
  • Basidiomycota / metabolism*
  • Disease Resistance*
  • Flax / genetics
  • Flax / metabolism*
  • Flax / microbiology
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Mutagenesis, Site-Directed
  • Plant Diseases*
  • Plant Proteins / genetics
  • Plant Proteins / metabolism*

Substances

  • Fungal Proteins
  • Plant Proteins