Cytochrome aa3 of Rhodobacter sphaeroides as a model for mitochondrial cytochrome c oxidase. The coxII/coxIII operon codes for structural and assembly proteins homologous to those in yeast.
The coxII/coxIII operon of Rhodobacter sphaeroides cytochrome c oxidase has been sequenced and characterized by insertional inactivation/complementation analysis. The organization of the genes in this locus (coxII.orf1.orf3.coxIII) is the same as that of the equivalent operon of Paracoccus denitrificans (ctaC.ctaB.ctaG.ctaE), but unlike that of other bacteria whose cytochrome oxidase genes have been characterized so far. The predicted amino acid sequence homology with eukaryotic oxidases is also higher for Rb. sphaeroides (and P. denitrificans) than for other bacterial versions of the enzyme. The inactivation of coxII results in loss of the characteristic cytochrome oxidase spectrum from membranes of the mutant strain. Full recovery requires introduction into the bacterium of the complete operon containing coxII.orf1.orf3.coxIII; partial complementation yielding a spectrally altered enzyme is achieved with a plasmid containing coxII or coxII.orf1.orf3. These results indicate that the peptides ORF1, ORF3, and COXIII are all required for assembly of native cytochrome c oxidase, suggesting an oxidase-specific assembly or chaperonin function for the ORFs in Rb. sphaeroides similar to that observed for the homologous gene products in yeast, COX10 and COX11.
Cao
J
J
Department of Biochemistry, Michigan State University, East Lansing 48824.
Hosler
J
J
Shapleigh
J
J
Revzin
A
A
Ferguson-Miller
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D003576:Electron Transport Complex IV / Q000235:genetics*; D005798:Genes, Bacterial; D005800:Genes, Fungal; D008322:Mammals; D008928:Mitochondria / Q000201:enzymology*; D008954:Models, Biological; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D009876:Operon; D010957:Plasmids; D015183:Restriction Mapping; D012242:Rhodobacter sphaeroides / Q000201:enzymology* / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics*; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D003576:Electron Transport Complex IV