title: Danger-associated peptide signaling in Arabidopsis requires clathrin.
abstract: The Arabidopsis thaliana endogenous elicitor peptides (AtPeps) are released into the apoplast after cellular damage caused by pathogens or wounding to induce innate immunity by direct binding to the membrane-localized leucine-rich repeat receptor kinases, PEP RECEPTOR1 (PEPR1) and PEPR2. Although the PEPR-mediated signaling components and responses have been studied extensively, the contributions of the subcellular localization and dynamics of the active PEPRs remain largely unknown. We used live-cell imaging of the fluorescently labeled and bioactive pep1 to visualize the intracellular behavior of the PEPRs in the Arabidopsis root meristem. We found that AtPep1 decorated the plasma membrane (PM) in a receptor-dependent manner and cointernalized with PEPRs. Trafficking of the AtPep1-PEPR1 complexes to the vacuole required neither the trans-Golgi network/early endosome (TGN/EE)-localized vacuolar H(+)-ATPase activity nor the function of the brefeldin A-sensitive ADP-ribosylation factor-guanine exchange factors (ARF-GEFs). In addition, AtPep1 and different TGN/EE markers colocalized only rarely, implying that the intracellular route of this receptor-ligand pair is largely independent of the TGN/EE. Inducible overexpression of the Arabidopsis clathrin coat disassembly factor, Auxilin2, which inhibits clathrin-mediated endocytosis (CME), impaired the AtPep1-PEPR1 internalization and compromised AtPep1-mediated responses. Our results show that clathrin function at the PM is required to induce plant defense responses, likely through CME of cell surface-located signaling components.
authors.affiliation: Department of Plant Systems Biology, VIB, 9052 Ghent, Belgium; Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Ghent, Belgium; Laboratory of Molecular and Plant Biology, Department of Genetics, College of Agriculture, University of São Paulo, Piracicaba, SP 13418-900, Brazil; Agricultural Engineering Program, University of the Amazon, Florencia 180002622, Colombia;
authors.forename: Fausto Andres
authors.initials: FA
authors.lastname: Ortiz-Morea
authors.affiliation: Department of Plant Systems Biology, VIB, 9052 Ghent, Belgium; Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Ghent, Belgium;
authors.forename: Daniel V
authors.initials: DV
authors.lastname: Savatin
authors.affiliation: Department of Plant Systems Biology, VIB, 9052 Ghent, Belgium; Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Ghent, Belgium;
authors.forename: Wim
authors.initials: W
authors.lastname: Dejonghe
authors.affiliation: Department of Plant Systems Biology, VIB, 9052 Ghent, Belgium; Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Ghent, Belgium;
authors.forename: Rahul
authors.initials: R
authors.lastname: Kumar
authors.affiliation: Department of Plant Systems Biology, VIB, 9052 Ghent, Belgium; Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Ghent, Belgium;
authors.forename: Yu
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Institute of Science and Technology Austria, 3400 Klosterneuburg, Austria;
authors.forename: Maciej
authors.initials: M
authors.lastname: Adamowski
authors.affiliation: Laboratory for Organic and Biomimetic Chemistry, Department of Organic Chemistry, Ghent University, 9000 Gent, Belgium.
authors.forename: Jos
authors.initials: J
authors.lastname: Van den Begin
authors.affiliation: Laboratory of Molecular and Plant Biology, Department of Genetics, College of Agriculture, University of São Paulo, Piracicaba, SP 13418-900, Brazil;
authors.forename: Keini
authors.initials: K
authors.lastname: Dressano
authors.affiliation: Department of Plant Systems Biology, VIB, 9052 Ghent, Belgium; Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Ghent, Belgium;
authors.forename: Guilherme
authors.initials: G
authors.lastname: Pereira de Oliveira
authors.affiliation: Department of Plant Systems Biology, VIB, 9052 Ghent, Belgium; Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Ghent, Belgium;
authors.forename: Xiuyang
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Department of Plant Systems Biology, VIB, 9052 Ghent, Belgium; Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Ghent, Belgium;
authors.forename: Qing
authors.initials: Q
authors.lastname: Lu
authors.affiliation: Laboratory for Organic and Biomimetic Chemistry, Department of Organic Chemistry, Ghent University, 9000 Gent, Belgium.
authors.forename: Annemieke
authors.initials: A
authors.lastname: Madder
authors.affiliation: Institute of Science and Technology Austria, 3400 Klosterneuburg, Austria;
authors.forename: Jiří
authors.initials: J
authors.lastname: Friml
authors.affiliation: Laboratory of Molecular and Plant Biology, Department of Genetics, College of Agriculture, University of São Paulo, Piracicaba, SP 13418-900, Brazil;
authors.forename: Daniel
authors.initials: D
authors.lastname: Scherer de Moura
authors.affiliation: Department of Plant Systems Biology, VIB, 9052 Ghent, Belgium; Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Ghent, Belgium; eurus@psb.vib-ugent.be.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Russinova
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism*; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018519:Meristem / Q000166:cytology / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D008954:Models, Biological; D010455:Peptides / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D012235:Rhodamines / Q000378:metabolism; D015398:Signal Transduction; D013347:Subcellular Fractions / Q000378:metabolism; D025262:Vacuolar Proton-Translocating ATPases / Q000378:metabolism; D021601:trans-Golgi Network / Q000378:metabolism
chemical_list: C114557:5-carboxytetramethylrhodamine succinimidyl ester; D029681:Arabidopsis Proteins; D002966:Clathrin; C512592:PEPR1 protein, Arabidopsis; D010455:Peptides; D011956:Receptors, Cell Surface; D012235:Rhodamines; D049452:Green Fluorescent Proteins; C582179:PEPR2 protein, Arabidopsis; D017346:Protein Serine-Threonine Kinases; D020928:Mitogen-Activated Protein Kinases; D025262:Vacuolar Proton-Translocating ATPases
keywords: Arabidopsis; PEPR; clathrin; endocytosis; endogenous peptides