title: Caenorhabditis elegans expressing the Saccharomyces cerevisiae NADH alternative dehydrogenase Ndi1p, as a tool to identify new genes involved in complex I related diseases.
abstract: Isolated complex I deficiencies are one of the most commonly observed biochemical features in patients suffering from mitochondrial disorders. In the majority of these clinical cases the molecular bases of the diseases remain unknown suggesting the involvement of unidentified factors that are critical for complex I function. The Saccharomyces cerevisiae NDI1 gene, encoding the mitochondrial internal NADH dehydrogenase was previously shown to complement a complex I deficient strain in Caenorhabditis elegans with notable improvements in reproduction and whole organism respiration. These features indicate that Ndi1p can functionally integrate the respiratory chain, allowing complex I deficiency complementation. Taking into account the Ndi1p ability to bypass complex I, we evaluate the possibility to extend the range of defects/mutations causing complex I deficiencies that can be alleviated by NDI1 expression. We report here that NDI1 expressing animals unexpectedly exhibit a slightly shortened lifespan, a reduction in the progeny, and a depletion of the mitochondrial genome. However, Ndi1p is expressed and targeted to the mitochondria as a functional protein that confers rotenone resistance to those animals without affecting their respiration rate and ATP content. We show that the severe embryonic lethality level caused by the RNAi knockdowns of complex I structural subunit encoding genes (e.g., NDUFV1, NDUFS1, NDUFS6, NDUFS8, or GRIM-19 human orthologs) in wild type animals is significantly reduced in the Ndi1p expressing worm. All together these results open up the perspective to identify new genes involved in complex I function, assembly, or regulation by screening an RNAi library of genes leading to embryonic lethality that should be rescued by NDI1 expression.
authors.affiliation: I2BC, Institute for Integrative Biology of the Cell, CEA, CNRS, Université Paris-Sud Orsay, France.
authors.forename: Raynald
authors.initials: R
authors.lastname: Cossard
authors.affiliation: I2BC, Institute for Integrative Biology of the Cell, CEA, CNRS, Université Paris-Sud Orsay, France.
authors.forename: Michela
authors.initials: M
authors.lastname: Esposito
authors.affiliation: I2BC, Institute for Integrative Biology of the Cell, CEA, CNRS, Université Paris-Sud Orsay, France.
authors.forename: Carole H
authors.initials: CH
authors.lastname: Sellem
authors.affiliation: I2BC, Institute for Integrative Biology of the Cell, CEA, CNRS, Université Paris-Sud Orsay, France.
authors.forename: Laras
authors.initials: L
authors.lastname: Pitayu
authors.affiliation: I2BC, Institute for Integrative Biology of the Cell, CEA, CNRS, Université Paris-Sud Orsay, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Vasnier
authors.affiliation: I2BC, Institute for Integrative Biology of the Cell, CEA, CNRS, Université Paris-Sud Orsay, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Delahodde
authors.affiliation: I2BC, Institute for Integrative Biology of the Cell, CEA, CNRS, Université Paris-Sud Orsay, France.
authors.forename: Emmanuel P
authors.initials: EP
authors.lastname: Dassa
keywords: Caenorhabditis elegans; Ndi1p; RNAi screening; complex I; embryonic lethality