title: Rapid screening for nuclear genes mutations in isolated respiratory chain complex I defects.
abstract: Complex I or reduced nicotinamide adenine dinucleotide (NADH): ubiquinone oxydoreductase deficiency is the most common cause of respiratory chain defects. Molecular bases of complex I deficiencies are rarely identified because of the dual genetic origin of this multi-enzymatic complex (nuclear DNA and mitochondrial DNA) and the lack of phenotype-genotype correlation. We used a rapid method to screen patients with isolated complex I deficiencies for nuclear genes mutations by Surveyor nuclease digestion of cDNAs. Eight complex I nuclear genes, among the most frequently mutated (NDUFS1, NDUFS2, NDUFS3, NDUFS4, NDUFS7, NDUFS8, NDUFV1 and NDUFV2), were studied in 22 cDNA fragments spanning their coding sequences in 8 patients with a biochemically proved complex I deficiency. Single nucleotide polymorphisms and missense mutations were detected in 18.7% of the cDNA fragments by Surveyor nuclease treatment. Molecular defects were detected in 3 patients. Surveyor nuclease screening is a reliable method for genotyping nuclear complex I deficiencies, easy to interpret, and limits the number of sequence reactions. Its use will enhance the possibility of prenatal diagnosis and help us for a better understanding of complex I molecular defects.
authors.affiliation: Laboratoire de Biochimie, APHP Hôpital de Bicêtre, 78, rue du Général Leclerc, 94275 Le Kremlin-Bicêtre Cedex, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Pagniez-Mammeri
authors.forename: Anne
authors.initials: A
authors.lastname: Lombes
authors.forename: Michèle
authors.initials: M
authors.lastname: Brivet
authors.forename: Hélène
authors.initials: H
authors.lastname: Ogier-de Baulny
authors.forename: Pierre
authors.initials: P
authors.lastname: Landrieu
authors.forename: Alain
authors.initials: A
authors.lastname: Legrand
authors.forename: Abdelhamid
authors.initials: A
authors.lastname: Slama
mesh_terms: D002467:Cell Nucleus / Q000235:genetics*; D002675:Child, Preschool; D018076:DNA, Complementary / Q000235:genetics; D003851:Deoxyribonucleases / Q000378:metabolism; D042967:Electron Transport Complex I / Q000172:deficiency* / Q000235:genetics*; D005820:Genetic Testing; D006801:Humans; D009154:Mutation / Q000235:genetics*; D010084:Oxidation-Reduction; D019289:Pyruvic Acid / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D019289:Pyruvic Acid; D003851:Deoxyribonucleases; D042967:Electron Transport Complex I