Deconstructing a molecular switch; structure-function analysis of the CC-NB-LRR proteins Rx1 and Gpa2

E.J. Slootweg, L.N. Spiridon, O. Caldararu, W. Tameling, H. Pomp, J. Roosien, J. Bakker, G. Smant, M. Joosten, A.J. Petrescu, A. Goverse

Research output: Chapter in Book/Report/Conference proceedingAbstract

Abstract

Many plant and animal immune receptors have a modular nucleotide-binding-leucine-rich repeat (NB-LRR) architecture in which a nucleotide-binding switch domain, NB-ARC, is tethered to a coiled coil (CC) and a LRR sensor domain. The cooperation between the CC, switch and sensor domains, which regulates the activation of these proteins, is poorly understood. Through targeted mutagenesis, interaction studies and structural modelling we attempt to investigate the way the domains of the potato resistance proteins Rx1 or Gpa2 work together. We demonstrated that the correct cooperation between the CC-NB-ARC and the LRR is focussed on a region in the ARC2 subdomain of the NB-ARC and the N-terminal repeats of the LRR. A mismatch leads either to autoactivation or loss-of-function. Complementary charged surface patches in the ARC2 and LRR are key determinants of the physical interaction between the domains, but do not appear to be involved in signalling. Furthermore alanine-substitution of specific aromatic residues in the CC domain shows that distinct surfaces are required for the interaction of the CC with the NBARC – LRR or with the protein RanGAP2. The CC domain of Rx1 consists of 4 helices and mutagenesis of the hydrophobic residues required for their interaction revealed that distinct parts of the CC are involved in either the cell death or PVX resistance signaling by Rx1. Currently we try to characterise how the binding of RanGAP2 and the recognition of specific elicitors affects these interdomain interactions and subcellular localisation of the resistance protein in the cell.
Original languageEnglish
Title of host publicationBook of abstracts of 2nd Annual Conference of the SUSTAIN Action, COST FA 1208 pathogen-informed strategies for sustainable broad-spectrum crop resistance
Pages24-24
Publication statusPublished - 2014
EventThe second Annual Conference of the SUSTAIN Action, Zakopane, Poland -
Duration: 15 Oct 201417 Oct 2014

Conference

ConferenceThe second Annual Conference of the SUSTAIN Action, Zakopane, Poland
Period15/10/1417/10/14

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