title: A cell biological view of the siderophore pyochelin iron uptake pathway in Pseudomonas aeruginosa.
abstract: Pyochelin (PCH) is a siderophore produced and secreted by Pseudomonas aeruginosa for iron capture. Using (55) Fe uptake and binding assays, we showed that PCH-Fe uptake in P. aeruginosa involves, in addition to the highly studied outer membrane transporter FptA, the inner membrane permease FptX, which recognizes PCH-(55) Fe with an affinity of 0.6 ± 0.2 nM and transports the ferri-siderophore complex from the periplasm into the cytoplasm: fptX deletion inhibited (55) Fe accumulation in the bacterial cytoplasm. Chromosomal replacement was used to generate P. aeruginosa strains producing fluorescent fusions with FptX, PchR (an AraC regulator), PchA (the first enzyme involved in the PCH biosynthesis) and PchE (a non-ribosomic peptide-synthetase involved in a further step). Fluorescence imaging and cellular fractionation showed a uniform repartition of FptX in the inner membrane. PchA and PchE were found in the cytoplasm, associated to the inner membrane all over the bacteria and also concentrated at the bacterial poles. PchE clustering at the bacterial poles was dependent on PchA expression, but on the opposite PchA clustering and membrane association was PchE-independent. PchA and PchE cellular organization suggests the existence of a siderosome for PCH biosynthesis as previously proposed for pyoverdine biosynthesis (another siderophore produced by P. aeruginosa).
authors.affiliation: UMR 7242, Université de Strasbourg-CNRS, ESBS, Blvd Sébastien Brant, Strasbourg, Illkirch, F-67413, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Cunrath
authors.forename: Véronique
authors.initials: V
authors.lastname: Gasser
authors.forename: Françoise
authors.initials: F
authors.lastname: Hoegy
authors.forename: Cornelia
authors.initials: C
authors.lastname: Reimmann
authors.forename: Laurent
authors.initials: L
authors.lastname: Guillon
authors.forename: Isabelle J
authors.initials: IJ
authors.lastname: Schalk
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D001692:Biological Transport; D007501:Iron / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000378:metabolism; D010636:Phenols / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D017262:Siderophores / Q000378:metabolism*; D013844:Thiazoles / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; D010636:Phenols; D017262:Siderophores; D013844:Thiazoles; C025316:pyochelin; D007501:Iron