title: Defining the pathogenesis of the human Atp12p W94R mutation using a Saccharomyces cerevisiae yeast model.
abstract: Studies in yeast have shown that a deficiency in Atp12p prevents assembly of the extrinsic domain (F(1)) of complex V and renders cells unable to make ATP through oxidative phosphorylation. De Meirleir et al. (De Meirleir, L., Seneca, S., Lissens, W., De Clercq, I., Eyskens, F., Gerlo, E., Smet, J., and Van Coster, R. (2004) J. Med. Genet. 41, 120-124) have reported that a homozygous missense mutation in the gene for human Atp12p (HuAtp12p), which replaces Trp-94 with Arg, was linked to the death of a 14-month-old patient. We have investigated the impact of the pathogenic W94R mutation on Atp12p structure/function. Plasmid-borne wild type human Atp12p rescues the respiratory defect of a yeast ATP12 deletion mutant (Deltaatp12). The W94R mutation alters the protein at the most highly conserved position in the Pfam sequence and renders HuAtp12p insoluble in the background of Deltaatp12. In contrast, the yeast protein harboring the corresponding mutation, ScAtp12p(W103R), is soluble in the background of Deltaatp12 but not in the background of Deltaatp12Deltafmc1, a strain that also lacks Fmc1p. Fmc1p is a yeast mitochondrial protein not found in higher eukaryotes. Tryptophan 94 (human) or 103 (yeast) is located in a positively charged region of Atp12p, and hence its mutation to arginine does not alter significantly the electrostatic properties of the protein. Instead, we provide evidence that the primary effect of the substitution is on the dynamic properties of Atp12p.
authors.affiliation: From the Center for Genetics, UZ Brussel, Vrije Universiteit Brussel, Brussels B-1050, Belgium.
authors.forename: Ann
authors.initials: A
authors.lastname: Meulemans
authors.affiliation: From the Center for Genetics, UZ Brussel, Vrije Universiteit Brussel, Brussels B-1050, Belgium.
authors.forename: Sara
authors.initials: S
authors.lastname: Seneca
authors.affiliation: the Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, Michigan 48201.
authors.forename: Thomas
authors.initials: T
authors.lastname: Pribyl
authors.affiliation: the Departments of Pediatrics, Division of Pediatric Neurology and Metabolism, Ghent University Hospital, Ghent 9000, Belgium.
authors.forename: Joel
authors.initials: J
authors.lastname: Smet
authors.affiliation: From the Center for Genetics, UZ Brussel, Vrije Universiteit Brussel, Brussels B-1050, Belgium.
authors.forename: Valerie
authors.initials: V
authors.lastname: Alderweirldt
authors.affiliation: Departments of Pathology, Division of Pediatric Neurology and Metabolism, Ghent University Hospital, Ghent 9000, Belgium, and.
authors.forename: Anouk
authors.initials: A
authors.lastname: Waeytens
authors.affiliation: From the Center for Genetics, UZ Brussel, Vrije Universiteit Brussel, Brussels B-1050, Belgium.
authors.forename: Willy
authors.initials: W
authors.lastname: Lissens
authors.affiliation: the Departments of Pediatrics, Division of Pediatric Neurology and Metabolism, Ghent University Hospital, Ghent 9000, Belgium.
authors.forename: Rudy
authors.initials: R
authors.lastname: Van Coster
authors.affiliation: Department of Pediatric Neurology, UZ Brussel, Vrije Universiteit Brussel, Brussels B-1050, Belgium.
authors.forename: Linda
authors.initials: L
authors.lastname: De Meirleir
authors.affiliation: the Institut de Biochimie et Génétique Cellulaires CNRS/Bordeaux 2 University, Bordeaux Cedex 33077, France.
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: di Rago
authors.affiliation: the Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, Michigan 48201.
authors.forename: Domenico L
authors.initials: DL
authors.lastname: Gatti
authors.affiliation: the Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, Michigan 48201. Electronic address: sackerm@med.wayne.edu.
authors.forename: Sharon H
authors.initials: SH
authors.lastname: Ackerman
mesh_terms: D019943:Amino Acid Substitution; D001120:Arginine / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002478:Cells, Cultured; D018833:Chaperonins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004579:Electron Transport / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure; D005816:Genetic Complementation Test; D006801:Humans; D008854:Microscopy, Electron; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism; D025261:Mitochondrial Proton-Translocating ATPases; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D011487:Protein Conformation; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D012995:Solubility; D055672:Static Electricity; D014364:Tryptophan / Q000235:genetics / Q000378:metabolism
chemical_list: C110382:ATP12 protein, S cerevisiae; C504819:Fmc1 protein, S cerevisiae; D024101:Mitochondrial Proteins; D018832:Molecular Chaperones; D029701:Saccharomyces cerevisiae Proteins; D014364:Tryptophan; D001120:Arginine; D018833:Chaperonins; C484671:ATPAF2 protein, human; D025261:Mitochondrial Proton-Translocating ATPases; D006180:Proton-Translocating ATPases