title: Rapid Oligo-Galacturonide Induced Changes in Protein Phosphorylation in Arabidopsis.
abstract: The wall-associated kinases (WAKs)(1)are receptor protein kinases that bind to long polymers of cross-linked pectin in the cell wall. These plasma-membrane-associated protein kinases also bind soluble pectin fragments called oligo-galacturonides (OGs) released from the wall after pathogen attack and damage. WAKs are required for cell expansion during development but bind water soluble OGs generated from walls with a higher affinity than the wall-associated polysaccharides. OGs activate a WAK-dependent, distinct stress-like response pathway to help plants resist pathogen attack. In this report, a quantitative mass-spectrometric-based phosphoproteomic analysis was used to identify Arabidopsis cellular events rapidly induced by OGsin planta Using N(14/)N(15)isotopicin vivometabolic labeling, we screened 1,000 phosphoproteins for rapid OG-induced changes and found 50 proteins with increased phosphorylation, while there were none that decreased significantly. Seven of the phosphosites within these proteins overlap with those altered by another signaling molecule plants use to indicate the presence of pathogens (the bacterial "elicitor" peptide Flg22), indicating distinct but overlapping pathways activated by these two types of chemicals. Genetic analysis of genes encoding 10 OG-specific and two Flg22/OG-induced phosphoproteins reveals that null mutations in eight proteins compromise the OG response. These phosphorylated proteins with genetic evidence supporting their role in the OG response include two cytoplasmic kinases, two membrane-associated scaffold proteins, a phospholipase C, a CDPK, an unknown cadmium response protein, and a motor protein. Null mutants in two proteins, the putative scaffold protein REM1.3, and a cytoplasmic receptor like kinase ROG2, enhance and suppress, respectively, a dominantWAKallele. Altogether, the results of these chemical and genetic experiments reveal the identity of several phosphorylated proteins involved in the kinase/phosphatase-mediated signaling pathway initiated by cell wall changes.
authors.affiliation: From the ‡Biology Department, Bowdoin College, Brunswick, ME, 04011; bkohorn@bowdoin.edu.
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Kohorn
authors.affiliation: From the ‡Biology Department, Bowdoin College, Brunswick, ME, 04011;
authors.forename: Divya
authors.initials: D
authors.lastname: Hoon
authors.affiliation: §Department of Biochemistry, University of Wisconsin, Madison, WI 53706.
authors.forename: Benjamin B
authors.initials: BB
authors.lastname: Minkoff
authors.affiliation: §Department of Biochemistry, University of Wisconsin, Madison, WI 53706.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Sussman
authors.affiliation: From the ‡Biology Department, Bowdoin College, Brunswick, ME, 04011;
authors.forename: Susan L
authors.initials: SL
authors.lastname: Kohorn
mesh_terms: D017360:Arabidopsis / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D002473:Cell Wall / Q000187:drug effects / Q000378:metabolism; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D007553:Isotope Labeling; D013058:Mass Spectrometry / Q000379:methods*; D009154:Mutation; D010368:Pectins / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction
chemical_list: D029681:Arabidopsis Proteins; D010368:Pectins; D011494:Protein Kinases
references.citation: Mol Plant. 2008 May;1(3):423-45
references.pmid: 19825551
references.citation: Plant J. 2009 Dec;60(6):974-82
references.pmid: 19737363
references.citation: Curr Biol. 2011 Oct 25;21(20):1720-6
references.pmid: 21982593
references.citation: Trends Plant Sci. 2010 Jul;15(7):363-9
references.pmid: 20483654
references.citation: BMC Plant Biol. 2007;7:16
references.pmid: 17397538
references.citation: Science. 2014 Jan 24;343(6169):408-11
references.pmid: 24458638
references.citation: Proteomics. 2010 Aug;10(16):3020-34
references.pmid: 20564262
references.citation: Proc Natl Acad Sci U S A. 2015 Jul 14;112(28):8780-5
references.pmid: 26124097
references.citation: Front Plant Sci. 2013 Mar 13;4:49
references.pmid: 23493833
references.citation: Plant Signal Behav. 2009 Aug;4(8):784-6
references.pmid: 19820315
references.citation: Plant Mol Biol. 2009 Feb;69(3):337-46
references.pmid: 19039666
references.citation: Proc Natl Acad Sci U S A. 2014 Oct 21;111(42):15261-6
references.pmid: 25288746
references.citation: Phytochemistry. 2006 Jun;67(11):1068-79
references.pmid: 16631829
references.citation: J Proteomics. 2011 Sep 6;74(10):1934-49
references.pmid: 21619950
references.citation: J Exp Bot. 2007;58(13):3719-30
references.pmid: 17928369
references.citation: Mol Cell Proteomics. 2007 Jul;6(7):1198-214
references.pmid: 17317660
references.citation: J Integr Plant Biol. 2013 Dec;55(12):1188-97
references.pmid: 23710768
references.citation: Plant Physiol. 2014 Feb;164(2):1093-107
references.pmid: 24367018
references.citation: Plant Physiol. 2005 Nov;139(3):1107-24
references.pmid: 16286450
references.citation: Plant Physiol. 2005 Jul;138(3):1334-46
references.pmid: 15951488
references.citation: Plant Physiol. 2003 Dec;133(4):1732-46
references.pmid: 14576286
references.citation: J Biol Chem. 2014 Jul 4;289(27):18978-86
references.pmid: 24855660
references.citation: Curr Biol. 2007 Jun 5;17(11):922-31
references.pmid: 17540573
references.citation: Plant Physiol. 2002 Jun;129(2):455-9
references.pmid: 12068092
references.citation: Curr Biol. 2009 Aug 11;19(15):1327-31
references.pmid: 19646876
references.citation: Plant Physiol. 2010 Feb;152(2):787-96
references.pmid: 19965966
references.citation: Proc Natl Acad Sci U S A. 2010 Sep 7;107(36):15986-91
references.pmid: 20733066
references.citation: Dev Cell. 2011 Dec 13;21(6):985-91
references.pmid: 22172668
references.citation: Curr Opin Plant Biol. 2008 Jun;11(3):321-8
references.pmid: 18395489
references.citation: Curr Protoc Bioinformatics. 2010 Mar;Chapter 13:Unit 13.12.1-11
references.pmid: 20205189
references.citation: Plant Physiol. 1983 Jul;72(3):864-9
references.pmid: 16663100
references.citation: Plant Physiol. 2000 Sep;124(1):31-8
references.pmid: 10982419
references.citation: Plant Mol Biol. 2001 Sep;47(1-2):9-27
references.pmid: 11554482
references.citation: Plant Cell. 2014 May 7;26(5):1848-1856
references.pmid: 24808052
references.citation: J Integr Plant Biol. 2013 Dec;55(12):1184-7
references.pmid: 24308569
references.citation: Genetics. 2005 Sep;171(1):305-21
references.pmid: 15965251
references.citation: Proc Natl Acad Sci U S A. 2010 May 18;107(20):9452-7
references.pmid: 20439716
references.citation: Front Plant Sci. 2012 May 07;3:86
references.pmid: 22639670
references.citation: Methods Mol Biol. 2014;1062:353-79
references.pmid: 24057376
references.citation: Mol Plant. 2012 Jul;5(4):841-51
references.pmid: 22155845
references.citation: Nat Genet. 2015 Feb;47(2):151-7
references.pmid: 25531751
references.citation: Front Plant Sci. 2012 Jun 06;3:121
references.pmid: 22685449
references.citation: Curr Opin Plant Biol. 2008 Jun;11(3):266-77
references.pmid: 18486536
references.citation: Plant Physiol. 2010 Jun;153(2):467-78
references.pmid: 20154095
references.citation: Mol Cell Proteomics. 2015 May;14(5):1169-82
references.pmid: 25693798
references.citation: Carbohydr Res. 2009 Sep 28;344(14):1879-900
references.pmid: 19616198
references.citation: Nat Methods. 2008 Apr;5(4):319-22
references.pmid: 18345006
references.citation: Front Plant Sci. 2012 Aug 03;3:177
references.pmid: 22876255
references.citation: J Biol Chem. 2012 Nov 16;287(47):39982-91
references.pmid: 23027878
references.citation: Front Plant Sci. 2012 May 08;3:88
references.pmid: 22639672
references.citation: Plant Cell. 2014 Mar;26(3):1018-35
references.pmid: 24681615
references.citation: Plant Cell. 1996 Dec;8(12):2265-76
references.pmid: 8989883
references.citation: Plant J. 2006 Apr;46(2):307-16
references.pmid: 16623892
references.citation: Curr Biol. 2012 Sep 25;22(18):1732-7
references.pmid: 22885061
references.citation: Proc Natl Acad Sci U S A. 2009 May 5;106(18):7648-53
references.pmid: 19383785
references.citation: Plant Physiol. 2014 May 7;165(3):1171-1187
references.pmid: 24808101
references.citation: Science. 2004 Dec 24;306(5705):2206-11
references.pmid: 15618507
references.citation: Plant Cell Physiol. 2005 Feb;46(2):268-78
references.pmid: 15769808
references.citation: Plant Physiol. 2010 Jun;153(2):384-95
references.pmid: 20427466