The secretome of the plant pathogenic bacterium Erwinia chrysanthemi.
Erwinia chrysanthemi causes soft-rot diseases of many plants by secreting a battery of enzymes which degrade the plant cell walls. We initiated a proteomic analysis to create a reference map of the E. chrysanthemi secretome. Extracellular proteins were isolated from E. chrysanthemi culture supernatants and resolved by two-dimensional electrophoresis. By analysis of mutants, Western blotting, and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) 55 spots representing 25 unique proteins were identified. In uninduced conditions, we identified spots corresponding to the cellulase Cel5, the proteases PrtA, PrtB, and PrtC, the flagellin FliC, and some intracellular proteins whose presence probably resulted from spontaneous cell lysis. We identified another secreted protein, AvrL, homologous to an avirulence protein of Xanthomonas campestris. After culture in conditions inducing pectinase production, i.e., in the presence of galacturonate and plant extract, we identified spots corresponding to the endopectate lyases PelA, PelB, PelC, PelD, PelE, PelI, PelL, and PelZ, the pectin acetylesterases PaeX and PaeY, the pectin methylesterase PemA, and the polygalacturonase PehX. In the presence of other inducing compounds, we detected the rhamnogalacturonate lyase RhiE and the esterase FaeD. Analysis of mutants, altered for one type of secretion system, was performed to determine the targets of each system. The type I system Prt was necessary for the secretion of three proteases. No proteins secreted by the type III Hrp system could be detected in E. chrysanthemi supernatants. In addition to the already known substrates (eleven pectinases and one cellulase), this analysis revealed that the type II Out system mediates secretion of the esterase FaeD and of the Avr-like protein AvrL.
Kazemi-Pour
Nasrin
N
Unit of Microbiology and Genetics, CNRS-UCB-INSA, La Doua Scientific Domain, Villeurbanne, France.
Condemine
Guy
G
Hugouvieux-Cotte-Pattat
Nicole
N
D001426:Bacterial Proteins / Q000737:chemistry*; D015153:Blotting, Western; D016974:Dickeya chrysanthemi / Q000235:genetics* / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005408:Flagellin / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005838:Genotype; D007477:Ions; D010641:Phenotype; D011096:Polygalacturonase / Q000737:chemistry*; D011133:Polysaccharide-Lyases / Q000737:chemistry; D011485:Protein Binding; D020543:Proteome; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D016959:Xanthomonas campestris / Q000378:metabolism
D001426:Bacterial Proteins; D007477:Ions; D020543:Proteome; D005408:Flagellin; D011096:Polygalacturonase; D011133:Polysaccharide-Lyases; C023911:pectate lyase