title: Functional characterization of novel mutations in the human cytochrome b gene.
abstract: The great variability of the human mitochondrial DNA (mtDNA) sequence induces many difficulties in the search for its deleterious mutations. We illustrate these pitfalls by the analysis of the cytochrome b gene of 21 patients affected with a mitochondrial disease. Eighteen different sequence variations were found, five of which were new mutations. Extensive analysis of the cytochrome b gene of 146 controls found 20 supplementary mutations, thus further demonstrating the high variability of the cytochrome b sequence. We fully evaluated the functional relevance of 36 of these 38 mutations using indirect criteria such as the nature of the mutation, its frequency in controls, or the phylogenetic conservation of the mutated amino acid. When appropriate, the mtDNA haplotype, the heteroplasmic state of the mutation, its tissue distribution or its familial transmission were also assessed. The molecular consequences of the mutations, which appeared possibly deleterious in that first step of evaluation, were evaluated on the complex III enzymological properties and protein composition using specific antibodies that we have generated against four of its subunits. Two original deleterious mutations were found in the group of seven patients with overt complex III defect. Both mutations (G15150A (W135X) and T15197C (S151P)) were heteroplasmic and restricted to muscle. They had significant consequences on the complex III structure. In contrast, only two homoplasmic missense mutations with dubious clinical relevance were found in the patients without overt complex III defect.
authors.affiliation: INSERM U523, Institut de Myologie, Hôpital de la Salpêtrière, 75651 Paris cedex 13, France.
authors.forename: F
authors.initials: F
authors.lastname: Legros
authors.forename: E
authors.initials: E
authors.lastname: Chatzoglou
authors.forename: P
authors.initials: P
authors.lastname: Frachon
authors.forename: H
authors.initials: H
authors.lastname: Ogier De Baulny
authors.forename: P
authors.initials: P
authors.lastname: Laforêt
authors.forename: C
authors.initials: C
authors.lastname: Jardel
authors.forename: C
authors.initials: C
authors.lastname: Godinot
authors.forename: A
authors.initials: A
authors.lastname: Lombès
mesh_terms: D019943:Amino Acid Substitution; D000968:Antimycin A / Q000031:analogs & derivatives* / Q000494:pharmacology; D015153:Blotting, Western; D003573:Cytochrome b Group / Q000235:genetics*; D004252:DNA Mutational Analysis; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics; D014450:Electron Transport Complex III / Q000187:drug effects / Q000378:metabolism; D005787:Gene Frequency; D014644:Genetic Variation; D006239:Haplotypes; D006801:Humans; D008689:Methacrylates; D017240:Mitochondrial Myopathies / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D017354:Point Mutation; D013844:Thiazoles / Q000494:pharmacology; D014451:Ubiquinone / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology
chemical_list: D003573:Cytochrome b Group; D004272:DNA, Mitochondrial; D008689:Methacrylates; D013844:Thiazoles; C032456:antimycin; D014451:Ubiquinone; C060262:2,3-dimethoxy-5-methyl-6-decyl-1,4-benzoquinone; D000968:Antimycin A; C030517:myxothiazol; D014450:Electron Transport Complex III