title: Mitochondrial frataxin interacts with ISD11 of the NFS1/ISCU complex and multiple mitochondrial chaperones.
abstract: The neurodegenerative disorder Friedreich's ataxia (FRDA) is caused by mutations in frataxin, a mitochondrial protein whose function remains controversial. Using co-immunoprecipitation and mass spectrometry we identified multiple interactors of mitochondrial frataxin in mammalian cells. One interactor was mortalin/GRP75, a homolog of the yeast ssq1 chaperone that integrates iron-sulfur clusters into imported mitochondrial proteins. Another interactor was ISD11, recently identified as a component of the eukaryotic complex Nfs1/ISCU, an essential component of iron-sulfur cluster biogenesis. Interactions between frataxin and ISD11, and frataxin and GRP75 were confirmed by co-immunoprecipitation experiments in both directions. Immunofluorescence analysis demonstrated that ISD11 co-localized with both frataxin and with mitochondria. The point mutations I154F and W155R in frataxin cause FRDA and are clustered to one surface of the protein, and these mutations decrease the interaction of frataxin with ISD11. The frataxin/ISD11 interaction was also decreased by the chelator EDTA, and was increased by supplementation with nickel but not other metal ions. Nickel supplementation rescued the defective interaction of mutant frataxin I154F and W155R with ISD11. Upon ISD11 depletion by siRNA in HEK293T cells, the amount of the Nfs1/ISCU protein complex declined, as did the activity of the iron-sulfur cluster enzyme aconitase, while the cellular iron content was increased, as seen in tissues from FRDA patients. Furthermore, ISD11 mRNA levels were decreased in FRDA patient cells. These data suggest that frataxin binds the iron-sulfur biogenesis Nfs1/ISCU complex through ISD11, that the interaction is nickel-dependent, and that multiple consequences of frataxin deficiency are duplicated by ISD11 deficiency.
authors.affiliation: VM:Molecular Biosciences, 1311 Haring Hall, Davis, CA 95616, USA.
authors.forename: Yuxi
authors.initials: Y
authors.lastname: Shan
authors.forename: Eleonora
authors.initials: E
authors.lastname: Napoli
authors.forename: Gino
authors.initials: G
authors.lastname: Cortopassi
mesh_terms: D000154:Aconitate Hydratase / Q000235:genetics; D000818:Animals; D019556:COS Cells; D013437:Carbon-Sulfur Lyases / Q000378:metabolism*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D005621:Friedreich Ataxia / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D007501:Iron / Q000378:metabolism; D033862:Iron-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D007506:Iron-Sulfur Proteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000378:metabolism*; D008954:Models, Biological; D018832:Molecular Chaperones / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000378:metabolism; D009532:Nickel / Q000502:physiology; D011485:Protein Binding; D021381:Protein Transport; D000096923:Frataxin
chemical_list: D000154:Aconitate Hydratase; D013437:Carbon-Sulfur Lyases; D018840:HSP70 Heat-Shock Proteins; D007501:Iron; D033862:Iron-Binding Proteins; D007506:Iron-Sulfur Proteins; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D018832:Molecular Chaperones; D046912:Multiprotein Complexes; D009532:Nickel; D000096923:Frataxin; C506160:ISCU protein, human; C018551:glucose-regulated proteins; C080352:mortalin; C513492:NFS1 protein, human