Isolation and characterization of QCR10, the nuclear gene encoding the 8.5-kDa subunit 10 of the Saccharomyces cerevisiae cytochrome bc1 complex.
We have cloned and sequenced QCR10, the nuclear gene encoding an 8.5-kDa protein proposed to be a subunit of the cytochrome bc1 complex of Saccharomyces cerevisiae. QCR10 includes a 231-base pair open reading frame capable of encoding a protein of 77 amino acids with a predicted molecular mass of 8492 Da. The codons for the amino-terminal methionine and alanine are separated from the remainder of the open reading frame by a 63-base pair intron that contains 5'-donor, 3'-acceptor, and TACTAAC sequences known to be necessary for splicing. The deduced amino acid sequence of the 8.5-kDa yeast protein is 28% identical to that of the 6.4-kDa subunit 11 from the bovine heart cytochrome bc1 complex, and the predicted secondary structures of the two proteins are very similar. Deletion of the chromosomal copy of QCR10 did not affect the growth of yeast on nonfermentable carbon sources. However, deletion of QCR10 synergistically contributes to the temperature-dependent phenotype resulting from deletion of QCR6, the gene for subunit 6 of the cytochrome bc1 complex. In addition, ubiquinol-cytochrome c oxidoreductase activity was reduced 40% in mitochondrial membranes from the QCR10 deletion strain, and the Rieske iron-sulfur protein was lost when the cytochrome bc1 complex lacking the 8.5-kDa protein was purified. We conclude that the 8.5-kDa protein encoded by QCR10 is a subunit of the yeast cytochrome bc1 complex and that the presence of this subunit during assembly is required for stable association of the iron-sulfur protein with the complex.
Brandt
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Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755-3844.
Uribe
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Schägger
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Trumpower
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D004271:DNA, Fungal; D014450:Electron Transport Complex III / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid
D004271:DNA, Fungal; D005656:Fungal Proteins; D009687:Nuclear Proteins; C087097:QCR10 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014450:Electron Transport Complex III