TABLE 1.

Distribution of dolichol and polyprenol in phosphorylated and neutral prenols of parental and mutant cells^a

Cell line	Prenol (%)
	Total		Phosphorylated		Neutral
	Phosphorylated	Neutral	Dolichol	Polyprenol	Dolichol	Polyprenol
WT	81 ± 4	20 ± 4	98 ± 1	2.5 ± 1	77 ± 16	23 ± 16
ConA 1-1	23 ± 19	77 ± 19	91 ± 7	9 ± 7	35 ± 9	65 ± 9

^aCells were incubated at 27°C with [³H] mevalonolactone for at least 36 h. Labeled lipids were sequentially extracted into chloroform-methanol (2:1) and then into chloroform-methanol-water (10:10:3) (for analysis of total prenol). Lipids extracted into chloroform-methanol (2:1) were resolved into phosphorylated and neutral fractions by chromatography on a DEAE-cellulose column (for analysis of dolichol and polyprenol). All fractions were saponified, and the alkali-resistant lipids were extracted into ether. Phosphorylated lipids were treated with acid phosphatase. Labeled lipids were fractionated by gel filtration chromatography, and the labeled prenols were analyzed by normal-phase silica HPLC. The amount of radioactivity was normalized for cell number and in the majority of experiments for the recovery of labeled internal standards. There were four determinations (three experiments) with WT cells and three determinations (three experiments) with ConA 1-1 cells.