2-hydroxyethylhydrazine as a potent inhibitor of phospholipid methylation in yeast

Abstract

The effect of 2-hydroxyethylhydrazine on the phosphatidylethanolamine methylation pathway in yeast was studied. 2-Hydroxyethylhydrazine inhibited the growth of cells. The concentration required for 50% inhibition was 66 microM. The growth rate decreased by 2-hydroxyethylhydrazine was restored by the addition of a low concentration of choline. Incorporation of radioactivity from L-[3-14C]serine, L-[methyl-14C]methionine and S-adenosyl-L-[methyl-14C]methionine into phosphatidylcholine was markedly reduced by 2-hydroxyethylhydrazine. The restoration of growth by choline was not due to the reversal of the inhibition, but to the formation of phosphatidylcholine via the CDPcholine pathway. Thus, the site of action of 2-hydroxyethylhydrazine in vivo was the phosphatidylethanolamine methylation pathway. Experiments with methylation mutants indicated that all three steps of methylation were sensitive to 2-hydroxyethylhydrazine. 2-Hydroxyethylhydrazine was shown to inhibit the methyltransferase after it had become chemically or metabolically transformed in cells. 2-Hydroxyethylhydrazine-resistant mutants were obtained and were found to have a defect in choline transport activity. Genetic data indicated that the uptake of 2-hydroxyethylhydrazine into cells is mediated by the choline transport system.