title: Both human ferredoxins 1 and 2 and ferredoxin reductase are important for iron-sulfur cluster biogenesis.
abstract: Ferredoxins are iron-sulfur proteins that have been studied for decades because of their role in facilitating the monooxygenase reactions catalyzed by p450 enzymes. More recently, studies in bacteria and yeast have demonstrated important roles for ferredoxin and ferredoxin reductase in iron-sulfur cluster assembly. The human genome contains two homologous ferredoxins, ferredoxin 1 (FDX1) and ferredoxin 2 (FDX2--formerly known as ferredoxin 1L). More recently, the roles of these two human ferredoxins in iron-sulfur cluster assembly were assessed, and it was concluded that FDX1 was important solely for its interaction with p450 enzymes to synthesize mitochondrial steroid precursors, whereas FDX2 was used for synthesis of iron-sulfur clusters, but not steroidogenesis. To further assess the role of the FDX-FDXR system in mammalian iron-sulfur cluster biogenesis, we performed siRNA studies on FDX1 and FDX2, on several human cell lines, using oligonucleotides identical to those previously used, along with new oligonucleotides that specifically targeted each gene. We concluded that both FDX1 and FDX2 were important in iron-sulfur cluster biogenesis. Loss of FDX1 activity disrupted activity of iron-sulfur cluster enzymes and cellular iron homeostasis, causing mitochondrial iron overload and cytosolic iron depletion. Moreover, knockdown of the sole human ferredoxin reductase, FDXR, diminished iron-sulfur cluster assembly and caused mitochondrial iron overload in conjunction with cytosolic depletion. Our studies suggest that interference with any of the three related genes, FDX1, FDX2 or FDXR, disrupts iron-sulfur cluster assembly and maintenance of normal cytosolic and mitochondrial iron homeostasis.
authors.affiliation: National Institute of Child Health and Human Development, Molecular Medicine Program, Bethesda, Maryland 20892, USA.
authors.forename: Yanbo
authors.initials: Y
authors.lastname: Shi
authors.forename: Manik
authors.initials: M
authors.lastname: Ghosh
authors.forename: Gennadiy
authors.initials: G
authors.lastname: Kovtunovych
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Crooks
authors.forename: Tracey A
authors.initials: TA
authors.lastname: Rouault
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D003600:Cytosol / Q000201:enzymology; D042967:Electron Transport Complex I / Q000378:metabolism; D005288:Ferredoxins / Q000235:genetics / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006418:Heme / Q000172:deficiency; D006801:Humans; D007501:Iron / Q000378:metabolism; D035941:Iron Regulatory Protein 1 / Q000378:metabolism; D035942:Iron Regulatory Protein 2 / Q000378:metabolism; D007506:Iron-Sulfur Proteins / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D005810:Multigene Family; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D016415:Sequence Alignment
chemical_list: D005288:Ferredoxins; D007506:Iron-Sulfur Proteins; D006418:Heme; D007501:Iron; D010088:Oxidoreductases; D035941:Iron Regulatory Protein 1; D035942:Iron Regulatory Protein 2; D042967:Electron Transport Complex I
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