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A novel allele in the Arabidopsis thaliana MACPF protein CAD1 results in deregulated immune signaling

View ORCID ProfileDanalyn R. Holmes, View ORCID ProfileMelissa Bredow, View ORCID ProfileKathrin Thor, Sydney A. Pascetta, View ORCID ProfileIrina Sementchoukova, Kristen R. Siegel, View ORCID ProfileCyril Zipfel, View ORCID ProfileJacqueline Monaghan
doi: https://doi.org/10.1101/2021.01.28.428637
Danalyn R. Holmes
*Biology Department, Queen’s University, Biological Sciences Complex, 116 Barrie St, K7L 3N6 Kingston, Ontario, Canada
1Center for Plant Molecular Biology, University of Tuebingen, Auf der Morgenstelle 32, D-72076 Tuebingen, Germany
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Melissa Bredow
*Biology Department, Queen’s University, Biological Sciences Complex, 116 Barrie St, K7L 3N6 Kingston, Ontario, Canada
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  • ORCID record for Melissa Bredow
Kathrin Thor
†The Sainsbury Laboratory, University of East Anglia, Norwich Research Park, NR4 7UH Norwich, Norfolk, United Kingdom
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Sydney A. Pascetta
*Biology Department, Queen’s University, Biological Sciences Complex, 116 Barrie St, K7L 3N6 Kingston, Ontario, Canada
2Department of Molecular and Cellular Biology, University of Guelph, Summerlee Science Complex, N1G 2W1, Guelph, Canada
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Irina Sementchoukova
*Biology Department, Queen’s University, Biological Sciences Complex, 116 Barrie St, K7L 3N6 Kingston, Ontario, Canada
3Department of Biochemistry and Biomedical Sciences, McMaster University, Michael DeGroote Centre for Learning and Discovery, L8S 4L8, Hamilton, Canada
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Kristen R. Siegel
*Biology Department, Queen’s University, Biological Sciences Complex, 116 Barrie St, K7L 3N6 Kingston, Ontario, Canada
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Cyril Zipfel
†The Sainsbury Laboratory, University of East Anglia, Norwich Research Park, NR4 7UH Norwich, Norfolk, United Kingdom
‡Department of Plant and Microbial Biology and Zurich-Basel Plant Science Center, University of Zurich, Zollikerstrasse 107, CH-8008 Zurich, Switzerland
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  • For correspondence: jacqueline.monaghan@queensu.ca cyril.zipfel@botinst.uzh.ch
Jacqueline Monaghan
*Biology Department, Queen’s University, Biological Sciences Complex, 116 Barrie St, K7L 3N6 Kingston, Ontario, Canada
†The Sainsbury Laboratory, University of East Anglia, Norwich Research Park, NR4 7UH Norwich, Norfolk, United Kingdom
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  • For correspondence: jacqueline.monaghan@queensu.ca cyril.zipfel@botinst.uzh.ch
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Abstract

Immune recognition in plants is governed by two major classes of receptors: pattern recognition receptors (PRRs) and nucleotide-binding leucine-rich repeat receptors (NLRs). Located at the cell surface, PRRs bind extracellular ligands originating from microbes (indicative of ‘non-self’) or damaged plant cells (indicative of ‘infected-self’), and trigger signaling cascades to protect against infection. Located intracellularly, NLRs sense pathogen-induced physiological changes and trigger localized cell death and systemic resistance. Immune responses are under tight regulation in order to maintain homeostasis and promote plant health. In a forward-genetic screen to identify regulators of PRR-mediated immune signaling, we identified a novel allele of the membrane-attack complex and perforin (MACPF)-motif containing protein CONSTITUTIVE ACTIVE DEFENSE 1 (CAD1) resulting from a missense mutation in a conserved N-terminal cysteine. We show that cad1-5 mutants display deregulated immune signaling and symptoms of autoimmunity dependent on the lipase-like protein ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1), suggesting that CAD1 integrity is monitored by the plant immune system. We further demonstrate that CAD1 localizes to both the cytosol and plasma membrane using confocal microscopy and subcellular fractionation. Our results offer new insights into immune homeostasis and provide tools to further decipher the intriguing role of MACPF proteins in plants.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted January 28, 2021.
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A novel allele in the Arabidopsis thaliana MACPF protein CAD1 results in deregulated immune signaling
Danalyn R. Holmes, Melissa Bredow, Kathrin Thor, Sydney A. Pascetta, Irina Sementchoukova, Kristen R. Siegel, Cyril Zipfel, Jacqueline Monaghan
bioRxiv 2021.01.28.428637; doi: https://doi.org/10.1101/2021.01.28.428637
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A novel allele in the Arabidopsis thaliana MACPF protein CAD1 results in deregulated immune signaling
Danalyn R. Holmes, Melissa Bredow, Kathrin Thor, Sydney A. Pascetta, Irina Sementchoukova, Kristen R. Siegel, Cyril Zipfel, Jacqueline Monaghan
bioRxiv 2021.01.28.428637; doi: https://doi.org/10.1101/2021.01.28.428637

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