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bioRxiv · 10.1101/592824

Structure-function analysis of ZAR1 immune receptor reveals key molecular interactions for activity

Baudin, M.·Schreiber, K. J.·Martin, E. C.·Petrescu, A. J.·Lewis, J.
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Abstract

NLR (Nucleotide-binding [NB] Leucine-rich repeat [LRR] Receptor) proteins are critical for inducing immune responses in response to pathogen proteins, and must be tightly regulated to prevent spurious activation in the absence of a pathogen. The ZAR1 NLR recognizes diverse effector proteins from Pseudomonas syringae, including HopZ1a, and Xanthomonas species. Receptor-like cytoplasmic kinases (RLCKs) such as ZED1, interact with ZAR1 and provide specificity for different effector proteins, such as HopZ1a. We previously developed a transient expression system in Nicotiana benthamiana, that allowed us to demonstrate ZAR1 function is conserved from the Brassicaceae to the Solanaceae. Here, we combined structural modeling of ZAR1, with molecular and functional assays in our transient system, to show that multiple intramolecular and intermolecular interactions regulate ZAR1 activity. We identified new determinants required for the formation of the ZARCC dimer and its activity. Lastly, we characterized new intramolecular interactions between ZAR1 subdomains that participate in keeping ZAR1 immune complexes inactive. This work identifies molecular constraints on immune receptor function and activation.\n\nOne sentence-summaryStructure-informed analyses reveal multiple finely-tuned intramolecular interactions that regulate the activity of the immune receptor ZAR1.\n\nFundingResearch on plant immunity in the Lewis laboratory was supported by the USDA ARS 2030-21000-046-00D and 2030-21000-050-00D (JDL), and the NSF Directorate for Biological Sciences IOS-1557661 (JDL). ECM and AJP acknowledge financial support from UEFISCDI grant PN-III-ID-PCE-2016-0650 and the Romanian Academy programs 1 & 2 of IBAR.

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Baudin, M., Schreiber, K. J., Martin, E. C., Petrescu, A. J., Lewis, J.. 2019-03-29. Structure-function analysis of ZAR1 immune receptor reveals key molecular interactions for activity. https://doi.org/10.1101/592824

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