title: PchC thioesterase optimizes nonribosomal biosynthesis of the peptide siderophore pyochelin in Pseudomonas aeruginosa.
abstract: In Pseudomonas aeruginosa, the antibiotic dihydroaeruginoate (Dha) and the siderophore pyochelin are produced from salicylate and cysteine by a thiotemplate mechanism involving the peptide synthetases PchE and PchF. A thioesterase encoded by the pchC gene was found to be necessary for maximal production of both Dha and pyochelin, but it was not required for Dha release from PchE and could not replace the thioesterase function specified by the C-terminal domain of PchF. In vitro, 2-aminobutyrate, a cysteine analog, was adenylated by purified PchE and PchF proteins. In vivo, this analog strongly interfered with Dha and pyochelin formation in a pchC deletion mutant but affected production of these metabolites only slightly in the wild type. Exogenously supplied cysteine overcame the negative effect of a pchC mutation to a large extent, whereas addition of salicylate did not. These data are in agreement with a role for PchC as an editing enzyme that removes wrongly charged molecules from the peptidyl carrier protein domains of PchE and PchF.
authors.affiliation: Département de Microbiologie Fondamentale, Bâtiment de Biologie, Université de Lausanne, Lausanne, Switzerland. Cornelia.Reimmann@imf.unil.ch
authors.forename: Cornelia
authors.initials: C
authors.lastname: Reimmann
authors.forename: Hiten M
authors.initials: HM
authors.lastname: Patel
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Walsh
authors.forename: Dieter
authors.initials: D
authors.lastname: Haas
mesh_terms: D000613:Aminobutyrates / Q000494:pharmacology; D001426:Bacterial Proteins / Q000502:physiology*; D003545:Cysteine / Q000494:pharmacology; D010636:Phenols / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D013844:Thiazoles
chemical_list: D000613:Aminobutyrates; D001426:Bacterial Proteins; C103415:PchC protein, Pseudomonas aeruginosa; D010636:Phenols; D013844:Thiazoles; C025316:pyochelin; C012223:alpha-aminobutyric acid; D003545:Cysteine