A study of the mechanism of selenite-induced hypomethylated DNA and differentiation of Friend erythroleukemic cells

Abstract

Sodium selenite is a good inducer of hemoglobin production in Friend erythroleukemic cells (FELC). At a concentration of 20 microM 70-80% of the cells produce hemoglobin and the DNA is hypomethylated. What is the mechanism for sodium selenite alteration of the DNA methylation pattern? Experiments with methionine adenosyltransferase (the enzyme that synthesizes adenosylmethionine) showed little effect of selenite on the activity of this enzyme in vitro or in vivo. Therefore, FELC are able to synthesize S-adenosylmethionine in the presence of sodium selenite. When sodium selenite was added to an in vitro assay for DNA methylase, the enzyme was non-competitively inhibited by 80% at 20 microM selenite with a Ki of 6 microM. DNA methylase isolated from control and selenite-treated FELC was purified through a DEAE-Sephacel column and no difference in activity was found. In the presence of selenite, DNA methylase is very sensitive to selenite inhibition, but removal of the selenite restores activity. However, DNA synthesized by FELC grown in the presence of selenite (no DNA methylase activity) was found to be hypomethylated. These results suggest that DNA methylase activity is inhibited in FELC grown in the presence of sodium selenite.