title: A Cysteine-Rich Protein Kinase Associates with a Membrane Immune Complex and the Cysteine Residues Are Required for Cell Death.
abstract: Membrane-localized proteins perceive and respond to biotic and abiotic stresses. We performed quantitative proteomics on plasma membrane-enriched samples from Arabidopsis (Arabidopsis thaliana) treated with bacterial flagellin. We identified multiple receptor-like protein kinases changing in abundance, including cysteine (Cys)-rich receptor-like kinases (CRKs) that are up-regulated upon the perception of flagellin. CRKs possess extracellular Cys-rich domains and constitute a gene family consisting of 46 members in Arabidopsis. The single transfer DNA insertion lines CRK28 and CRK29, two CRKs induced in response to flagellin perception, did not exhibit robust alterations in immune responses. In contrast, silencing of multiple bacterial flagellin-induced CRKs resulted in enhanced susceptibility to pathogenic Pseudomonas syringae, indicating functional redundancy in this large gene family. Enhanced expression of CRK28 in Arabidopsis increased disease resistance to P. syringae Expression of CRK28 in Nicotiana benthamiana induced cell death, which required intact extracellular Cys residues and a conserved kinase active site. CRK28-mediated cell death required the common receptor-like protein kinase coreceptor BAK1. CRK28 associated with BAK1 as well as the activated FLAGELLIN-SENSING2 (FLS2) immune receptor complex. CRK28 self-associated as well as associated with the closely related CRK29. These data support a model where Arabidopsis CRKs are synthesized upon pathogen perception, associate with the FLS2 complex, and coordinately act to enhance plant immune responses.
authors.affiliation: Department of Plant Pathology (K.A.Y., J.M.E., A.Y.C., J.Y.F., G.C., J.S.R.) and Genome Center Proteomics Core Facility (B.P.), University of California, Davis, California 95616; and.|Department of Biochemistry and Biophysics and Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas 77843 (B.F.).
authors.forename: Koste A
authors.identifier: 0000-0001-5579-8150
authors.initials: KA
authors.lastname: Yadeta
authors.affiliation: Department of Plant Pathology (K.A.Y., J.M.E., A.Y.C., J.Y.F., G.C., J.S.R.) and Genome Center Proteomics Core Facility (B.P.), University of California, Davis, California 95616; and.|Department of Biochemistry and Biophysics and Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas 77843 (B.F.).
authors.forename: James M
authors.initials: JM
authors.lastname: Elmore
authors.affiliation: Department of Plant Pathology (K.A.Y., J.M.E., A.Y.C., J.Y.F., G.C., J.S.R.) and Genome Center Proteomics Core Facility (B.P.), University of California, Davis, California 95616; and.|Department of Biochemistry and Biophysics and Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas 77843 (B.F.).
authors.forename: Athena Y
authors.initials: AY
authors.lastname: Creer
authors.affiliation: Department of Plant Pathology (K.A.Y., J.M.E., A.Y.C., J.Y.F., G.C., J.S.R.) and Genome Center Proteomics Core Facility (B.P.), University of California, Davis, California 95616; and.|Department of Biochemistry and Biophysics and Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas 77843 (B.F.).
authors.forename: Baomin
authors.initials: B
authors.lastname: Feng
authors.affiliation: Department of Plant Pathology (K.A.Y., J.M.E., A.Y.C., J.Y.F., G.C., J.S.R.) and Genome Center Proteomics Core Facility (B.P.), University of California, Davis, California 95616; and.|Department of Biochemistry and Biophysics and Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas 77843 (B.F.).
authors.forename: Jessica Y
authors.initials: JY
authors.lastname: Franco
authors.affiliation: Department of Plant Pathology (K.A.Y., J.M.E., A.Y.C., J.Y.F., G.C., J.S.R.) and Genome Center Proteomics Core Facility (B.P.), University of California, Davis, California 95616; and.|Department of Biochemistry and Biophysics and Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas 77843 (B.F.).
authors.forename: Jose Sebastian
authors.identifier: 0000-0002-3871-3706
authors.initials: JS
authors.lastname: Rufian
authors.affiliation: Department of Plant Pathology (K.A.Y., J.M.E., A.Y.C., J.Y.F., G.C., J.S.R.) and Genome Center Proteomics Core Facility (B.P.), University of California, Davis, California 95616; and.|Department of Biochemistry and Biophysics and Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas 77843 (B.F.).
authors.forename: Ping
authors.initials: P
authors.lastname: He
authors.affiliation: Department of Plant Pathology (K.A.Y., J.M.E., A.Y.C., J.Y.F., G.C., J.S.R.) and Genome Center Proteomics Core Facility (B.P.), University of California, Davis, California 95616; and.|Department of Biochemistry and Biophysics and Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas 77843 (B.F.).
authors.forename: Brett
authors.initials: B
authors.lastname: Phinney
authors.affiliation: Department of Plant Pathology (K.A.Y., J.M.E., A.Y.C., J.Y.F., G.C., J.S.R.) and Genome Center Proteomics Core Facility (B.P.), University of California, Davis, California 95616; and glcoaker@ucdavis.edu.|Department of Biochemistry and Biophysics and Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas 77843 (B.F.) glcoaker@ucdavis.edu.
authors.forename: Gitta
authors.identifier: 0000-0003-0899-2449
authors.initials: G
authors.lastname: Coaker
mesh_terms: D017360:Arabidopsis / Q000166:cytology* / Q000276:immunology / Q000382:microbiology; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D020134:Catalytic Domain; D016923:Cell Death / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology; D003545:Cysteine / Q000378:metabolism*; D005408:Flagellin / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D030821:Plants, Genetically Modified; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D044224:Pseudomonas syringae / Q000472:pathogenicity; D017382:Reactive Oxygen Species / Q000378:metabolism; D014026:Nicotiana / Q000166:cytology / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins; D017382:Reactive Oxygen Species; D005408:Flagellin; D011494:Protein Kinases; C463660:BAK1 protein, Arabidopsis; C411853:FLS2 protein, Arabidopsis; C000624998:CRK28 protein, Arabidopsis; C000624999:CRK29 protein, Arabidopsis; D017346:Protein Serine-Threonine Kinases; D003545:Cysteine