Complementation of Saccharomyces cerevisiae coq7 mutants by mitochondrial targeting of the Escherichia coli UbiF polypeptide: two functions of yeast Coq7 polypeptide in coenzyme Q biosynthesis.
Coenzyme Q (ubiquinone or Q) functions in the respiratory electron transport chain and serves as a lipophilic antioxidant. In the budding yeast Saccharomyces cerevisiae, Q biosynthesis requires nine Coq proteins (Coq1-Coq9). Previous work suggests both an enzymatic activity and a structural role for the yeast Coq7 protein. To define the functional roles of yeast Coq7p we test whether Escherichia coli ubiF can functionally substitute for yeast COQ7. The ubiF gene encodes a flavin-dependent monooxygenase that shares no homology to the Coq7 protein and is required for the final monooxygenase step of Q biosynthesis in E. coli. The ubiF gene expressed at low copy restores growth of a coq7 point mutant (E194K) on medium containing a non-fermentable carbon source, but fails to rescue a coq7 null mutant. However, expression of ubiF from a multicopy vector restores growth and Q synthesis for both mutants, although with a higher efficiency in the point mutant. We attribute the more efficient rescue of the coq7 point mutant to higher steady state levels of the Coq3, Coq4, and Coq6 proteins and to the presence of demethoxyubiquinone, the substrate of UbiF. Coq7p co-migrates with the Coq3 and Coq4 polypeptides as a high molecular mass complex. Here we show that addition of Q to the growth media also stabilizes the Coq3 and Coq4 polypeptides in the coq7 null mutant. The data suggest that Coq7p, and the lipid quinones (demethoxyubiquinone and Q) function to stabilize other Coq polypeptides.
Tran
UyenPhuong C
UC
Department of Chemistry and Biochemistry, and Molecular Biology Institute, University of California-Los Angeles, 607 Charles E. Young Drive East, Los Angeles, CA 90095, USA.
Marbois
Beth
B
Gin
Peter
P
Gulmezian
Melissa
M
Jonassen
Tanya
T
Clarke
Catherine F
CF
D000595:Amino Acid Sequence; D004563:Electrochemistry; D029968:Escherichia coli Proteins / Q000235:genetics*; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D008055:Lipids; D008928:Mitochondria / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000235:genetics*; D008956:Models, Chemical; D008969:Molecular Sequence Data; D009154:Mutation; D017354:Point Mutation; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D014451:Ubiquinone / Q000737:chemistry*
D029968:Escherichia coli Proteins; D008055:Lipids; D014451:Ubiquinone; D006899:Mixed Function Oxygenases; C410160:UbiF protein, E coli