Subfamily C7 Raf-like kinases MRK1, RAF26, and RAF39 regulate immune homeostasis and stomatal opening in Arabidopsis thaliana

Gonçalves Dias, Márcia, Doss, Bassem, Rawat, Anamika, Siegel, Kristen R., Mahathanthrige, Tharika, Sklenar, Jan, Rodriguez Gallo, Maria Camila, Derbyshire, Paul, Dharmasena, Thakshila, Cameron, Emma, Uhrig, R. Glen, Zipfel, Cyril, Menke, Frank L. H. ORCID: https://orcid.org/0000-0003-2490-4824 and Monaghan, Jacqueline (2024) Subfamily C7 Raf-like kinases MRK1, RAF26, and RAF39 regulate immune homeostasis and stomatal opening in Arabidopsis thaliana. New Phytologist, 244 (6). pp. 2278-2294. ISSN 0028-646X

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Abstract

The calcium-dependent protein kinase CPK28 regulates several stress pathways in multiple plant species. Here, we aimed to discover CPK28-associated proteins in Arabidopsis thaliana. We used affinity-based proteomics and identified several potential CPK28 binding partners, including the C7 Raf-like kinases MRK1, RAF26, and RAF39. We used biochemistry, genetics, and physiological assays to gain insight into their function. We define redundant roles for these kinases in stomatal opening, immune-triggered reactive oxygen species (ROS) production, and resistance to a bacterial pathogen. We report that CPK28 associates with and trans-phosphorylates RAF26 and RAF39, and that MRK1, RAF26, and RAF39 are active kinases that localize to endomembranes. Although Raf-like kinases share some features with mitogen-activated protein kinase kinase kinases (MKKKs), we found that MRK1, RAF26, and RAF39 are unable to trans-phosphorylate any of the 10 Arabidopsis mitogen-activated protein kinase kinases (MKKs). Overall, our study suggests that C7 Raf-like kinases associate with and are phosphorylated by CPK28, function redundantly in stomatal opening and immunity, and possess substrate specificities distinct from canonical MKKKs.

Item Type: Article
Additional Information: Data availability statement: Any novel germplasm or clones described in this article will be made freely available upon request. The person responsible for sharing materials is the author of correspondence jacqueline.monaghan@queensu.ca. Proteomics data has been deposited to the ProteomeXchange Consortium via the PRIDE (Perez-Riverol et al., 2019; https://www.ebi.ac.uk/pride/) partner repository with the dataset identifiers PXD052803 and PXD055293. Funding Information: Biotechnology and Biological Sciences Research Council; Ontario Ministry of Research and Innovation. Grant Number: ER21-16-100; Natural Sciences and Engineering Research Council of Canada. Grant Numbers: RGPAS-492902-2016, RGPIN-2016-04787.
Uncontrolled Keywords: arabidopsis thaliana,c7 raf-like kinase,cpk28,immunity,mrk1,raf26,raf39,stomata,physiology,plant science ,/dk/atira/pure/subjectarea/asjc/1300/1314
Faculty \ School: Faculty of Science > The Sainsbury Laboratory
Faculty of Science > School of Biological Sciences
UEA Research Groups: Faculty of Science > Research Groups > Plant Sciences
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Depositing User: LivePure Connector
Date Deposited: 14 Jan 2025 00:59
Last Modified: 18 Jan 2025 01:08
URI: https://ueaeprints.uea.ac.uk/id/eprint/98197
DOI: 10.1111/nph.20198

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