Distal genes of the nuo operon of Rhodobacter capsulatus equivalent to the mitochondrial ND subunits are all essential for the biogenesis of the respiratory NADH-ubiquinone oxidoreductase.
Seven out of the 13 proteins encoded by the mitochondrial genome of mammals (peptides ND1 to ND6 plus ND4L) are subunits of the respiratory NADH-ubiquinone oxidoreductase (complex I). The function of these ND subunits is still poorly understood. We have used the NADH-ubiquinone oxidoreductase of Rhodobacter capsulatus as a model for the study of the function of these proteins. In this bacterium, the 14 genes encoding the NADH-ubiquinone oxidoreductase are clustered in the nuo operon. We report here on the biochemical and spectroscopic characterization of mutants individually disrupted in five nuo genes, equivalent to mitochondrial genes nd1, nd2, nd5, nd6 and nd4L. Disruption of any of these genes in R. capsulatus leads to the suppression of NADH dehydrogenase activity at the level of the bacterial membranes and to the disappearance of complex I-associated iron-sulphur clusters. Individual NUO subunits can still be immunodetected in the membranes of these mutants, but they do not form a functional subcomplex. In contrast to these observations, disruption of two ORFs (orf6 and orf7), also present in the distal part of the nuo operon, does not suppress NADH dehydrogenase activity or complex I-associated EPR signals, thus demonstrating that these ORFs are not essential for the biosynthesis of complex I.
Dupuis
A
A
Laboratoire de BioEnergétique Cellulaire et Pathologique, EA 2019 UJF, DBMS, CEA, Grenoble, France. dupuis@aup.ceng.cea.fr
Darrouzet
E
E
Duborjal
H
H
Pierrard
B
B
Chevallet
M
M
van Belzen
R
R
Albracht
S P
SP
Lunardi
J
J
D001426:Bacterial Proteins / Q000502:physiology; D004578:Electron Spin Resonance Spectroscopy; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D009154:Mutation; D009247:NADH, NADPH Oxidoreductases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D009876:Operon; D010957:Plasmids / Q000235:genetics; D016354:Rhodobacter capsulatus / Q000201:enzymology / Q000235:genetics*
D001426:Bacterial Proteins; D009247:NADH, NADPH Oxidoreductases