title: A single external enzyme confers alternative NADH:ubiquinone oxidoreductase activity in Yarrowia lipolytica.
abstract: NADH:ubiquinone oxidoreductases catalyse the first step within the diverse pathways of mitochondrial NADH oxidation. In addition to the energy-conserving form commonly called complex I, fungi and plants contain much simpler alternative NADH:ubiquinone oxido-reductases that catalyze the same reaction but do not translocate protons across the inner mitochondrial membrane. Little is known about the distribution and function of these enzymes. We have identified YLNDH2 as the only gene encoding an alternative NADH:ubiquinone oxidoreductase (NDH2) in the obligate aerobic yeast Yarrowia lipolytica. Cells carrying a deletion of YLNDH2 were fully viable; full inhibition by piericidin A indicated that complex I activity was the sole NADH:ubiquinone oxidoreductase activity left in the deletion strains. Studies with intact mitochondria revealed that NDH2 in Y. lipolytica is oriented towards the external face of the mitochondrial inner membrane. This is in contrast to the situation seen in Saccharomyces cerevisiae, Neurospora crassa and in green plants, where internal alternative NADH:ubiquinone oxidoreductases have been reported. Phylogenetic analysis of known NADH:ubiquinone oxidoreductases suggests that during evolution conversion of an ancestral external alternative NADH:ubiquinone oxidoreductase to an internal enzyme may have paved the way for the loss of complex I in fermenting yeasts like S. cerevisiae.
authors.affiliation: Institut für Biochemie I, Zentrum der Biologischen Chemie, Universitätsklinikum Frankfurt, D-60590 Frankfurt am Main, Germany.
authors.forename: S J
authors.initials: SJ
authors.lastname: Kerscher
authors.forename: J G
authors.initials: JG
authors.lastname: Okun
authors.forename: U
authors.initials: U
authors.lastname: Brandt
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D042967:Electron Transport Complex I; D017353:Gene Deletion; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D017510:Protein Folding; D004718:Saccharomycetales / Q000201:enzymology* / Q000235:genetics; D017386:Sequence Homology, Amino Acid
chemical_list: D017931:DNA Primers; D009247:NADH, NADPH Oxidoreductases; D042967:Electron Transport Complex I