title: Diiron centre mutations in Ciona intestinalis alternative oxidase abolish enzymatic activity and prevent rescue of cytochrome oxidase deficiency in flies.
abstract: The mitochondrial alternative oxidase, AOX, carries out the non proton-motive re-oxidation of ubiquinol by oxygen in lower eukaryotes, plants and some animals. Here we created a modified version of AOX from Ciona instestinalis, carrying mutations at conserved residues predicted to be required for chelation of the diiron prosthetic group. The modified protein was stably expressed in mammalian cells or flies, but lacked enzymatic activity and was unable to rescue the phenotypes of flies knocked down for a subunit of cytochrome oxidase. The mutated AOX transgene is thus a potentially useful tool in studies of the physiological effects of AOX expression.
authors.affiliation: BioMediTech and Tampere University Hospital, University of Tampere, FI-33014, Finland.
authors.forename: Ana
authors.initials: A
authors.lastname: Andjelković
authors.affiliation: BioMediTech and Tampere University Hospital, University of Tampere, FI-33014, Finland.|Departamento de Tecnologia, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista "Júlio de Mesquita Filho", 14884-900 Jaboticabal, SP, Brazil.
authors.forename: Marcos T
authors.initials: MT
authors.lastname: Oliveira
authors.affiliation: BioMediTech and Tampere University Hospital, University of Tampere, FI-33014, Finland.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Cannino
authors.affiliation: BioMediTech and Tampere University Hospital, University of Tampere, FI-33014, Finland.
authors.forename: Cagri
authors.initials: C
authors.lastname: Yalgin
authors.affiliation: BioMediTech and Tampere University Hospital, University of Tampere, FI-33014, Finland.|Institute of Biotechnology, University of Helsinki, FI-00014, Finland.
authors.forename: Praveen K
authors.initials: PK
authors.lastname: Dhandapani
authors.affiliation: BioMediTech and Tampere University Hospital, University of Tampere, FI-33014, Finland.
authors.forename: Eric
authors.initials: E
authors.lastname: Dufour
authors.affiliation: INSERM UMR 1141 and Université Paris 7, Faculté de Médecine Denis Diderot, Hôpital Robert Debré, 48, Boulevard Sérurier, 75019, Paris, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Rustin
authors.affiliation: BioMediTech and Tampere University Hospital, University of Tampere, FI-33014, Finland.|Institute of Biotechnology, University of Helsinki, FI-00014, Finland.
authors.forename: Marten
authors.initials: M
authors.lastname: Szibor
authors.affiliation: BioMediTech and Tampere University Hospital, University of Tampere, FI-33014, Finland.|Institute of Biotechnology, University of Helsinki, FI-00014, Finland.
authors.forename: Howard T
authors.initials: HT
authors.lastname: Jacobs
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D002938:Ciona intestinalis / Q000201:enzymology* / Q000235:genetics; D029721:Drosophila Proteins / Q000172:deficiency / Q000235:genetics; D004331:Drosophila melanogaster / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D003576:Electron Transport Complex IV / Q000235:genetics / Q000378:metabolism*; D005260:Female; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006801:Humans; D007501:Iron / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010101:Oxygen Consumption; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D000099188:Alternative Oxidase
chemical_list: D029721:Drosophila Proteins; D003576:Electron Transport Complex IV; D007501:Iron; D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D000099188:Alternative Oxidase