title: Quantitative proteomics reveals a dynamic association of proteins to detergent-resistant membranes upon elicitor signaling in tobacco.
abstract: A large body of evidence from the past decade supports the existence, in membrane from animal and yeast cells, of functional microdomains playing important roles in protein sorting, signal transduction, or infection by pathogens. In plants, as previously observed for animal microdomains, detergent-resistant fractions, enriched in sphingolipids and sterols, were isolated from plasma membrane. A characterization of their proteic content revealed their enrichment in proteins involved in signaling and response to biotic and abiotic stress and cell trafficking suggesting that these domains were likely to be involved in such physiological processes. In the present study, we used (14)N/(15)N metabolic labeling to compare, using a global quantitative proteomics approach, the content of tobacco detergent-resistant membranes extracted from cells treated or not with cryptogein, an elicitor of defense reaction. To analyze the data, we developed a software allowing an automatic quantification of the proteins identified. The results obtained indicate that, although the association to detergent-resistant membranes of most proteins remained unchanged upon cryptogein treatment, five proteins had their relative abundance modified. Four proteins related to cell trafficking (four dynamins) were less abundant in the detergent-resistant membrane fraction after cryptogein treatment, whereas one signaling protein (a 14-3-3 protein) was enriched. This analysis indicates that plant microdomains could, like their animal counterpart, play a role in the early signaling process underlying the setup of defense reaction. Furthermore proteins identified as differentially associated to tobacco detergent-resistant membranes after cryptogein challenge are involved in signaling and vesicular trafficking as already observed in similar studies performed in animal cells upon biological stimuli. This suggests that the ways by which the dynamic association of proteins to microdomains could participate in the regulation of the signaling process may be conserved between plant and animals.
authors.affiliation: Institut National de Recherche Agronomique, Unité Mixte de Recherche Plante Microbe Environnement 1088/CNRS 5184/Université de Bourgogne, 17 Rue Sully, BP 86510 F-21000 Dijon, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Stanislas
authors.forename: David
authors.initials: D
authors.lastname: Bouyssie
authors.forename: Michel
authors.initials: M
authors.lastname: Rossignol
authors.forename: Simona
authors.initials: S
authors.lastname: Vesa
authors.forename: Jérôme
authors.initials: J
authors.lastname: Fromentin
authors.forename: Johanne
authors.initials: J
authors.lastname: Morel
authors.forename: Carole
authors.initials: C
authors.lastname: Pichereaux
authors.forename: Bernard
authors.initials: B
authors.lastname: Monsarrat
authors.forename: Françoise
authors.initials: F
authors.lastname: Simon-Plas
mesh_terms: D020418:Algal Proteins / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology*; D005656:Fungal Proteins; D008163:Luminescent Measurements; D013058:Mass Spectrometry; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D040901:Proteomics / Q000379:methods*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013194:Staining and Labeling; D014026:Nicotiana / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000382:microbiology
chemical_list: D020418:Algal Proteins; D003902:Detergents; D005656:Fungal Proteins; D010455:Peptides; D010940:Plant Proteins; D017382:Reactive Oxygen Species; C061032:cryptogein protein, Phytophthora cryptogea