title: Siderophore-controlled iron assimilation in the enterobacterium Erwinia chrysanthemi: evidence for the involvement of bacterioferritin and the Suf iron-sulfur cluster assembly machinery.
abstract: The intracellular fate of iron acquired by bacteria during siderophore-mediated assimilation is poorly understood. We investigated this question in the pathogenic enterobacterium Erwinia chrysanthemi. This bacterium produces two siderophores, chrysobactin and achromobactin, during plant infection. We analyzed the distribution of iron into cytosolic proteins in bacterial cells supplied with 59Fe-chrysobactin using native gel electrophoresis. A parental strain and mutants deficient in bacterioferritin (bfr), miniferritin (dps), ferritin (ftnA), bacterioferredoxin (bfd), or iron-sulfur cluster assembly machinery (sufABCDSE) were studied. In the parental strain, we observed two rapidly 59Fe-labeled protein signals identified as bacterioferritin and an iron pool associated to the protein chain-elongation process. In the presence of increased 59Fe-chrysobactin concentrations, we detected mini-ferritin-bound iron. Iron incorporation into bacterioferritin was severely reduced in nonpolar sufA, sufB, sufD, sufS, and sufE mutants but not in a sufC background. Iron recycling from bacterioferritin did not occur in bfd and sufC mutants. Iron depletion caused a loss of aconitase activity, whereas ferric chrysobactin supplementation stimulated the production of active aconitase in parental cells and in bfr and bfd mutants. Aconitase activity in sufA, sufB, sufD, sufS, and sufE mutant strains was 10 times lower than that in parental cells. In the sufC mutant, it was twice as low as that in the parental strain. Defects observed in the mutants were not caused by altered ferric chrysobactin transport. Our data demonstrate a functional link between bacterioferritin, bacterioferredoxin, and the Suf protein machinery resulting in optimal bacterial growth and a balanced distribution of iron between essential metalloproteins.
authors.affiliation: Laboratoire Interactions Plantes-Pathogènes, Unité Mixte de Recherche 217, Institut National de la Recherche Agronomique, AgroParisTech, Université Paris 6, 75005 Paris, France. expert@agroparistech.fr
authors.forename: Dominique
authors.initials: D
authors.lastname: Expert
authors.forename: Aïda
authors.initials: A
authors.lastname: Boughammoura
authors.forename: Thierry
authors.initials: T
authors.lastname: Franza
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000502:physiology; D001692:Biological Transport; D003573:Cytochrome b Group / Q000737:chemistry* / Q000502:physiology; D016974:Dickeya chrysanthemi / Q000378:metabolism*; D004151:Dipeptides / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005293:Ferritins / Q000737:chemistry* / Q000502:physiology; D015964:Gene Expression Regulation, Bacterial; D007501:Iron / Q000737:chemistry / Q000378:metabolism*; D007506:Iron-Sulfur Proteins / Q000737:chemistry*; D008667:Metalloproteins / Q000737:chemistry; D008954:Models, Biological; D009154:Mutation; D010084:Oxidation-Reduction; D010957:Plasmids / Q000378:metabolism; D017262:Siderophores / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D003573:Cytochrome b Group; D004151:Dipeptides; D007506:Iron-Sulfur Proteins; D008667:Metalloproteins; D017262:Siderophores; C058543:chrysobactin; D005293:Ferritins; C018032:bacterioferritin; D007501:Iron