Ubiquinone biosynthesis in Saccharomyces cerevisiae: the molecular organization of O-methylase Coq3p depends on Abc1p/Coq8p.
Coenzyme Q is a redox-active lipid that functions as an electron carrier in the mitochondrial respiratory chain. Q-biosynthesis in Saccharomyces cerevisiae requires at least nine proteins (Coq1p-Coq9p). The molecular function of Coq8p is still unknown; however, lack of Q and the concomitant accumulation of the intermediate 3-hexaprenyl-4-hydroxybenzoic acid in the absence of Coq8p suggest an essential role in Q-biosynthesis. Localization studies identify Coq8p as a soluble mitochondrial protein, with characteristics of a protein of the matrix or associated with the inner mitochondrial membrane. Coq8p forms homomeric structure(s) as revealed by two-hybrid analysis and tandem affinity purification. Two-dimensional (2D)-Blue Native/sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis suggests that Coq8p - together with Coq2p and Coq10p - is predominantly associated with a complex of about 500 kDa, whereas Coq3p, Coq5p and Coq9p are mainly organized in a 1.3 MDa Q-biosynthesis complex that is not associated with the complex III and IV supracomplexes of the respiratory chain. Loss of Coq8p is accompanied by destabilization of Coq3p, but not of Coq9p from the 1.3 MDa Q-biosynthesis complex. This effect cannot be reversed by Q(6) supplementation. The detection of Coq3p isoforms by 2D-isoelectric focusing is in line with the proposed function of Coq8p as a kinase, with Coq3p as a target.
Tauche
Anja
A
Institute of Genetics, Dresden University of Technology, Dresden, Germany.
Krause-Buchholz
Udo
U
Rödel
Gerhard
G
D015966:Gene Expression Regulation, Fungal; D008780:Methyltransferases / Q000235:genetics* / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D010770:Phosphotransferases / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014451:Ubiquinone / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism
C099891:COQ8 protein, S cerevisiae; D024101:Mitochondrial Proteins; D029701:Saccharomyces cerevisiae Proteins; D014451:Ubiquinone; C030778:ubiquinone 8; D008780:Methyltransferases; C070696:3,4-dihydroxy-5-hexaprenylbenzoate methyltransferase; D010770:Phosphotransferases