Identification of proteins interacting with the tomato NB-LRR NRC1

D.J. Sueldo, T.W.H. Liebrand, Q. Liu, J.H. Vossen, M. Prins, M.H.A.J. Joosten, W.I.L. Tameling

Research output: Chapter in Book/Report/Conference proceedingAbstract

Abstract

Plant disease resistance (R) proteins confer immunnity to pathogens possessing the corresponding avirulence (Avr) proteins. Most R proteins localize intracellularly and only a few (e.g Cfs) are extracellular transmembrane proteins. Most intracellular R proteins belong to the nucleotide-binding and
leucine-rich repeat (NB-LRR) class. Activation of NB-LRRs is often associated with induction of the hypersensitive response (HR), a form of programmed cell death. NRC1 (NB-LRR required for HR-associated cell death-1) is an NB-LRR from tomato which participates in the signaling cascade required for
Cf-mediated resistance to Cladosporium fulvum. Silencing of NRC1 in Nicotiana benthamiana results in an impaired HR induced by extracellular and intracellular R proteins (Gabriëls et al. (2007), Plant J 50:14-28). To understand how NRC1
regulates the defense response induced by multiple R proteins we focus on the identification of interacting proteins. A thylakoid lumen protein was identified in a yeast two-hybrid screening. Additional yeast two-hybrid and
co-immunoprecipitation assays with in planta-expressed proteins, employing different domains of NRC1, are in progress. Silencing and overexpression of the thylakoid lumen protein should reveal whether it is required for NRC1 function.
Identification of more interactors will be carried out by affinity-purification of NRC1 expressed in planta followed by mass spectrometry analysis. The effect on affinity-tags on NRC1 activity was tested by fusing them to an autoactive
NRC1 mutant displaying an elicitor-independent HR. In most cases only a slight inhibition on NRC1 acivity was observed. With this research we intend to elucidate whether NRC1 is a true signaling hub for multiple R proteins, required to induce defense responses.
Original languageEnglish
Title of host publicationISMPMI International Congress abstracts, Quebec City, Canada, 19-23 July 2009
PublisherInternational Society for Molecular Plant-Microbe Interactions
Publication statusPublished - 2009
EventXIV International Congress on Molecular Plant-Microbe Interactions, Quebec City, Canada -
Duration: 19 Jul 200923 Jul 2009

Conference

ConferenceXIV International Congress on Molecular Plant-Microbe Interactions, Quebec City, Canada
Period19/07/0923/07/09

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