Cytochrome oxidase assembly does not require catalytically active cytochrome C.
Cytochrome c oxidase (COX), the terminal enzyme of the mitochondrial respiratory chain, catalyzes the transfer of electrons from reduced cytochrome c to molecular oxygen. COX assembly requires the coming together of nuclear- and mitochondrial-encoded subunits and the assistance of a large number of nuclear gene products acting at different stages of maturation of the enzyme. In Saccharomyces cerevisiae, expression of cytochrome c, encoded by CYC1 and CYC7, is required not only for electron transfer but also for COX assembly through a still unknown mechanism. We have attempted to distinguish between a functional and structural requirement of cytochrome c in COX assembly. A cyc1/cyc7 double null mutant strain was transformed with the cyc1-166 mutant gene (Schweingruber, M. E., Stewart, J. W., and Sherman, F. (1979) J. Biol. Chem. 254, 4132-4143) that expresses stable but catalytically inactive iso-1-cytochrome c. The COX content of the cyc1/cyc7 double mutant strain harboring non-functional iso-1-cytochrome c has been characterized spectrally, functionally, and immunochemically. The results of these studies demonstrate that cytochrome c plays a structural rather than functional role in assembly of cytochrome c oxidase. In addition to its requirement for COX assembly, cytochrome c also affects turnover of the enzyme. Mutants containing wild type apocytochrome c in mitochondria lack COX, suggesting that only the folded and mature protein is able to promote COX assembly.
Barrientos
Antoni
A
Department of Biological Sciences, Columbia University, New York, New York, 10027, USA.
Pierre
Danielle
D
Lee
Johnson
J
Tzagoloff
Alexander
A
D000483:Alleles; D001059:Apoproteins / Q000378:metabolism; D002384:Catalysis; D002455:Cell Division; D002467:Cell Nucleus / Q000378:metabolism; D003574:Cytochrome c Group / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D045304:Cytochromes c; D004247:DNA / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000737:chemistry* / Q000378:metabolism; D019286:Endopeptidase K / Q000378:metabolism; D005656:Fungal Proteins; D008928:Mitochondria / Q000378:metabolism; D009154:Mutation; D010100:Oxygen / Q000378:metabolism; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D017510:Protein Folding; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000378:metabolism; D013053:Spectrophotometry; D013696:Temperature; D013997:Time Factors
D001059:Apoproteins; D003574:Cytochrome c Group; D005656:Fungal Proteins; D045304:Cytochromes c; D004247:DNA; D003576:Electron Transport Complex IV; D019286:Endopeptidase K; D010100:Oxygen