The yeast complex I equivalent NADH dehydrogenase rescues pink1 mutants

• Published in: PLoS genetics Vol. 8; no. 1; p. e1002456
• Main Authors: Vilain, Sven, Esposito, Giovanni, Haddad, Dominik, Schaap, Onno, Dobreva, Mariya P, Vos, Melissa, Van Meensel, Stefanie, Morais, Vanessa A, De Strooper, Bart, Verstreken, Patrik
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 01.01.2012; Public Library of Science (PLoS)
• Subjects: Animals; Animals, Genetically Modified - genetics; Animals, Genetically Modified - metabolism; Biology; Ciona intestinalis; Ciona intestinalis - genetics; Cytoskeletal Proteins - genetics; Cytoskeletal Proteins - metabolism; Defects; Deoxyribonucleic acid; DNA; Drosophila; Drosophila melanogaster - genetics; Drosophila melanogaster - metabolism; Drosophila Proteins - genetics; Drosophila Proteins - metabolism; Electron transport; Electron Transport Complex I - genetics; Electron Transport Complex I - metabolism; Electron Transport Complex III - genetics; Electron Transport Complex III - metabolism; Electron Transport Complex IV - genetics; Electron Transport Complex IV - metabolism; Enzymes; Evacuations & rescues; Experiments; Gene Expression Regulation; Genetic aspects; Genetic engineering; Grants; GTP-Binding Proteins - genetics; GTP-Binding Proteins - metabolism; Humans; Insects; Male; Marine; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mitochondria; Mitochondria - genetics; Mitochondria - metabolism; Mitochondrial DNA; Mitochondrial Proteins - metabolism; Mutation; NADH dehydrogenase; Oxidoreductases - metabolism; Parkinson Disease - genetics; Parkinson's disease; Physiological aspects; Plant Proteins - metabolism; Protein-Serine-Threonine Kinases - genetics; Protein-Serine-Threonine Kinases - metabolism; Risk factors; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Ubiquitin-Protein Ligases - genetics; Ubiquitin-Protein Ligases - metabolism; Yeast; Belgium
• ISSN: 1553-7404; 1553-7390; 1553-7404
• DOI: 10.1371/journal.pgen.1002456

Pink1 is a mitochondrial kinase involved in Parkinson's disease, and loss of Pink1 function affects mitochondrial morphology via a pathway involving Parkin and components of the mitochondrial remodeling machinery. Pink1 loss also affects the enzymatic activity of isolated Complex I of the electron transport chain (ETC); however, the primary defect in pink1 mutants is unclear. We tested the hypothesis that ETC deficiency is upstream of other pink1-associated phenotypes. We expressed Saccaromyces cerevisiae Ndi1p, an enzyme that bypasses ETC Complex I, or sea squirt Ciona intestinalis AOX, an enzyme that bypasses ETC Complex III and IV, in pink1 mutant Drosophila and find that expression of Ndi1p, but not of AOX, rescues pink1-associated defects. Likewise, loss of function of subunits that encode for Complex I-associated proteins displays many of the pink1-associated phenotypes, and these defects are rescued by Ndi1p expression. Conversely, expression of Ndi1p fails to rescue any of the parkin mutant phenotypes. Additionally, unlike pink1 mutants, fly parkin mutants do not show reduced enzymatic activity of Complex I, indicating that Ndi1p acts downstream or parallel to Pink1, but upstream or independent of Parkin. Furthermore, while increasing mitochondrial fission or decreasing mitochondrial fusion rescues mitochondrial morphological defects in pink1 mutants, these manipulations fail to significantly rescue the reduced enzymatic activity of Complex I, indicating that functional defects observed at the level of Complex I enzymatic activity in pink1 mutant mitochondria do not arise from morphological defects. Our data indicate a central role for Complex I dysfunction in pink1-associated defects, and our genetic analyses with heterologous ETC enzymes suggest that Ndi1p-dependent NADH dehydrogenase activity largely acts downstream of, or in parallel to, Pink1 but upstream of Parkin and mitochondrial remodeling.