Relation of Mevalonate Synthesis to Mitochondrial Ubiquinone Content and Respiratory Function in Cultured Neuroblastoma Cells

The consequence of blocking the de novo synthesis of ubiquinone (coenzyme Q) on mitochondrial ubiquinone content and respiratory function was studied in cultured C1300 (Neuro 2A) murine neuroblastoma cells. Mevinolin, a competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, was used to suppress the synthesis of mevalonate, an essential precursor for the isoprenoid side chain of ubiquinone. At a concentration of 25 microM, mevinolin completely inhibited the incorporation of [3H]acetate into ubiquinone, isolated from cell extracts by two-dimensional thin-layer chromatography. Similar results were obtained when [14C]tyrosine was used as a precursor for the quinone ring. Through the use of reverse-phase thin-layer chromatography, it was established that the principal product of the ubiquinone pathway in murine neuroblastoma cells was ubiquinone-9. Inhibition of ubiquinone synthesis for 24h in cells cultured in the presence of 10% fetal calf serum (which contains 0.14 nmol of ubiquinone/ml of serum) resulted in a 40-57% decline in the concentration of ubiquinone in the mitochondria. However, the activities of succinate-cytochrome c reductase and succinate dehydrogenase in whole-cell homogenates or mitochondria were not inhibited. The state 3 and uncoupled rates of respiration, determined by polarographic measurements of oxygen consumption in homogenates and mitochondria, were elevated slightly in the mevinolin-treated cells. The data demonstrate that, although mevalonate synthesis is important for the maintenance of the intramitochondrial ubiquinone pool in cultured cells, major changes in the ubiquinone content of the mitochondria can occur in intact cells without perturbation of respiratory function. However, the coincidence of decreased mitochondrial ubiquinone concentration and the inhibition of cell cycling previously observed in mevinolin-treated cells (Maltese, W.A. (1984) Biochem. Biophys. Res. Commun. 120, 454-460) suggests that the availability of ubiquinone may play a role in the regulation of mitochondrial and cellular proliferation.