Molecular genetics of the mammalian NADH-ubiquinone oxidoreductase

A serendipitous observation led to the first characterization of a respiration-deficient Chinese hamster mutant cell line. It has guided the design of an enrichment scheme for the isolation of additional mutant cell lines. Several complementation groups were identified with mutations affecting complex I. The X-linked NDUFA1 gene encoding the MWFE protein represents one group. Several mutant alleles isolated independently are described that yield very low activities and demonstrate that the MWFE protein is essential for activity. A phylogenetic sequence analysis of this highly conserved protein has directed attention to species-specific differences that make the primate MWFE protein inactive in hamster cells. Based on such comparisons, mutant alleles made by site-directed mutagenesis were expressed in a null mutant and reduced complex I activities were observed, with the mutant protein assembled into the complex. These and other mutants promise to be valuable for structure-function analyses, especially in conjunction with a high-resolution structure to be expected in the future. The possibility for transgenic and knock-in mice as models for mitochondrial diseases is being explored.