FARNESOL IS UTILIZED FOR PROTEIN ISOPRENYLATION AND THE BIOSYNTHESIS OF CHOLESTEROL IN MAMMALIAN-CELLS

Evidence has been obtained indicating that free farnesol (F-OH) can be utilized for isoprenoid biosynthesis in mammalian cells. When rat C6 glial cells and an African green monkey kidney cell line (CV-1) were i ncubated with [H-3]F-OH, radioactivity was incorporated into cholester ol, ubiquinone (CoQ) and isoprenylated proteins. The incorporation of label from [H-3]F-OH into cholesterol in C6 and CV-1 cells was blocked by squalestatin 1 (SQ) which specifically inhibits the conversion of farnesyl pyrophosphate (F-P-P) to squalene. This result strongly sugge sts that cholesterol, and probably CoQ and protein, is metabolically l abeled via F-P-P. SDS-PAGE analysis of the delipidated protein fractio ns from C6 and CV-1 cells revealed several labeled polypeptides. Consi stent with these proteins being modified by isoprenylation of cysteine residues, Pronase E digestion released a major labeled product with t he chromatographic mobility of [H-3]farnesyl-cysteine (F-Cys). A diffe rent set of polypeptides was labeled when C6 and CV-1 cells were incub ated with [H-3]geranylgeraniol (GG-OH). Both sets of proteins appear t o be metabolically labeled by [H-3]mevalonolactone, and [H-3]-labeled F-Cys and geranylgeranyl-cysteine (GG-Cys) were liberated from these p roteins by Pronase E treatment. These cellular and biochemical studies indicate that F-OH can be used for isoprenoid biosynthesis and protei n isoprenylation in mammalian cells after being converted to F-P-P by phosphorylation reactions that remain to be elucidated. (C) 1995 Acade mic Press, Inc.