title: Mrs3p, Mrs4p, and frataxin provide iron for Fe-S cluster synthesis in mitochondria.
abstract: Yeast Mrs3p and Mrs4p are evolutionarily conserved mitochondrial carrier proteins that transport iron into mitochondria under some conditions. Yeast frataxin (Yfh1p), the homolog of the human protein implicated in Friedreich ataxia, is involved in iron homeostasis. However, its precise functions are controversial. Anaerobically grown triple mutant cells (Deltamrs3/4/Deltayfh1) displayed a severe growth defect corrected by in vivo iron supplementation. Because anaerobically grown cells do not synthesize heme, and they do not experience oxidative stress, this growth defect was most likely due to Fe-S cluster deficiency. Fe-S cluster formation was assessed in anaerobically grown cells shifted to air for a brief period. In isolated mitochondria, Fe-S clusters were detected on newly imported yeast ferredoxin precursor and on endogenous aconitase by means of [35S]cysteine labeling and native gel separation. New cluster formation was dependent on iron addition to mitochondria, and the iron concentration dependence was shifted dramatically upward in the Deltamrs3/4 mutant, indicating a role of Mrs3/4p in iron transport. The frataxin mutant strain lacked protein import capacity because of low mitochondrial membrane potential, although this was partially restored by growth in the presence of high iron. Under these conditions, a kinetic defect in new Fe-S cluster formation was still noted. Import of frataxin into frataxin-minus isolated mitochondria promptly corrected the Fe-S cluster assembly defect without further iron addition. These findings show that Mrs3/4p transports iron into mitochondria, whereas frataxin makes iron already within mitochondria available for Fe-S cluster synthesis.
authors.affiliation: Department of Medicine, Division of Hematology-Oncology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Elise R
authors.initials: ER
authors.lastname: Lyver
authors.forename: Simon A B
authors.initials: SA
authors.lastname: Knight
authors.forename: Debkumar
authors.initials: D
authors.lastname: Pain
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Lesuisse
authors.forename: Andrew
authors.initials: A
authors.lastname: Dancis
mesh_terms: D027682:Cation Transport Proteins / Q000378:metabolism / Q000502:physiology*; D005838:Genotype; D007501:Iron / Q000378:metabolism; D033862:Iron-Binding Proteins / Q000235:genetics / Q000502:physiology*; D007506:Iron-Sulfur Proteins / Q000737:chemistry*; D007700:Kinetics; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism; D051336:Mitochondrial Membranes / Q000378:metabolism; D024101:Mitochondrial Proteins; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism / Q000502:physiology*; D000096923:Frataxin
chemical_list: D027682:Cation Transport Proteins; D007501:Iron; D033862:Iron-Binding Proteins; D007506:Iron-Sulfur Proteins; D024101:Mitochondrial Proteins; D029701:Saccharomyces cerevisiae Proteins; D000096923:Frataxin; C066985:MRS3 protein, S cerevisiae; C066986:MRS4 protein, S cerevisiae