title: Frataxin and mitochondrial carrier proteins, Mrs3p and Mrs4p, cooperate in providing iron for heme synthesis.
abstract: Frataxin is a conserved mitochondrial protein implicated in cellular iron metabolism. Deletion of the yeast frataxin homolog (YFH1) was combined with deletions of MRS3 and MRS4, mitochondrial carrier proteins implicated in iron homeostasis. As previously reported, the Deltayfh1 mutant accumulated iron in mitochondria, whereas the triple mutant (DeltaDeltaDelta) did not. When wild-type, Deltamrs3/4, Deltayfh1, and DeltaDeltaDelta strains were incubated anaerobically, all strains were devoid of heme and protected from iron and oxygen toxicity. The cultures were then shifted to air for a short time (4-5 h) or a longer time (15 h), and the evolving mutant phenotypes were analyzed (heme-dependent growth, total heme, cytochromes, heme proteins, and iron levels). A picture emerges from these data of defective heme formation in the mutants, with a markedly more severe defect in the DeltaDeltaDelta than in the individual Deltamrs3/4 or Deltayfh1 mutants (a "synthetic" defect in the genetic sense). The defect(s) in heme formation could be traced to lack of iron. Using a real time assay of heme biosynthesis, porphyrin precursor and iron were presented to permeabilized cells, and the appearance and disappearance of fluorescent porphyrins were followed. The Mrs3/4p carriers were required for rapid iron transport into mitochondria for heme synthesis, whereas there was also evidence for an alternative slower system. A different role for Yfh1p was observed under conditions of low mitochondrial iron and aerobic growth (revealed in the DeltaDeltaDelta), acting to protect bioavailable iron within mitochondria and to facilitate its use for heme synthesis.
authors.affiliation: Department of Medicine, Division of Hematology-Oncology, University of Pennsylvania, Philadelphia, 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: Emmanuel
authors.initials: E
authors.lastname: Lesuisse
authors.forename: Andrew
authors.initials: A
authors.lastname: Dancis
mesh_terms: D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism*; D003433:Crosses, Genetic; D017353:Gene Deletion; D005800:Genes, Fungal; D006418:Heme / Q000096:biosynthesis*; D007501:Iron / Q000378:metabolism*; D033862:Iron-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D000096923:Frataxin
chemical_list: D027682:Cation Transport Proteins; D006418:Heme; D007501:Iron; D033862:Iron-Binding Proteins; D024101:Mitochondrial Proteins; D029701:Saccharomyces cerevisiae Proteins; D000096923:Frataxin; C066985:MRS3 protein, S cerevisiae; C066986:MRS4 protein, S cerevisiae