The relationship between intracellular sulfhydryl(SH) compounds and the kinetics of the inhibitory effect of selenite on cellular nucleic acid synthesis has been examined. In A549 cells, with a relatively high SH level, exposure to low concentrations of selenite caused inhibition even after short exposure times. In contrast, in VA cells, with a relatively low level of SH compounds, selenite had no significant effect at short exposure times, but inhibited significantly with longer exposures. Selenodicysteine, the product of the reaction of selenite with cysteine (an important intracellular SH compound), inhibited synthesis in both cell types at short exposure times. Exposure of cells to diethylmaleate, which decreased the level of intracellular SH compounds, reduced the inhibitory effect of a short exposure to selenite but did not affect a long exposure. These results indicate that the reaction of selenite with intracellular SH compounds may be a determining factor in the kinetics of its inhibitory effect on cellular DNA and RNA synthesis.