Trafficking of siderophore transporters in Saccharomyces cerevisiae and intracellular fate of ferrioxamine B conjugates.
We have studied the intracellular trafficking of Sit1 [ferrioxamine B (FOB) transporter] and Enb1 (enterobactin transporter) in Saccharomyces cerevisiae using green fluorescent protein (GFP) fusion proteins. Enb1 was constitutively targeted to the plasma membrane. Sit1 was essentially targeted to the vacuolar degradation pathway when synthesized in the absence of substrate. Massive plasma membrane sorting of Sit1 was induced by various siderophore substrates of Sit1, and by coprogen, which is not a substrate of Sit1. Thus, different siderophore transporters use different regulated trafficking processes. We also studied the fate of Sit1-mediated internalized siderophores. Ferrioxamine B was recovered in isolated vacuolar fractions, where it could be detected spectrophotometrically. Ferrioxamine B coupled to an inhibitor of mitochondrial protoporphyrinogen oxidase (acifluorfen) could not reach its target unless the cells were disrupted, confirming the tight compartmentalization of siderophores within cells. Ferrioxamine B coupled to a fluorescent moiety, FOB-nitrobenz-2-oxa-1,3-diazole, used as a Sit1-dependent iron source, accumulated in the vacuolar lumen even in mutants displaying a steady-state accumulation of Sit1 at the plasma membrane or in endosomal compartments. Thus, the fates of siderophore transporters and siderophores diverge early in the trafficking process.
Froissard
Marine
M
Laboratoire Trafic intracellulaire des protéines dans la levure, Département de biologie Cellulaire, Institut Jacques Monod, Unité Mixte de Recherche 7592 CNRS-Universités Paris 6 et 7, France.
Belgareh-Touzé
Naïma
N
Dias
Marylène
M
Buisson
Nicole
N
Camadro
Jean-Michel
JM
Haguenauer-Tsapis
Rosine
R
Lesuisse
Emmanuel
E
D001692:Biological Transport; D002176:Candida albicans / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D003676:Deferoxamine / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D005290:Ferric Compounds / Q000737:chemistry*; D005838:Genotype; D049452:Green Fluorescent Proteins / Q000378:metabolism; D020128:Inhibitory Concentration 50; D007501:Iron / Q000737:chemistry; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism; D008956:Models, Chemical; D021381:Protein Transport; D050768:Protoporphyrinogen Oxidase / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D017262:Siderophores / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
D005290:Ferric Compounds; D017262:Siderophores; D049452:Green Fluorescent Proteins; C002577:ferrioxamine B; D007501:Iron; D050768:Protoporphyrinogen Oxidase; D003676:Deferoxamine
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