Mitogenic stimulation of protein synthesis is accompanied by an increase in eIF-4E phosphorylation. The effect on protein synthesis by induction of differentiation is less well known. We treated P19 embryonal carcinoma cells with the differentiating agent retinoic acid and found that protein synthesis increased during the first hour of addition. However, the phosphorylation state, as well as the turnover of phosphate on eIF-4E, remained unchanged. Apparently, the change in protein synthesis after RA addition is regulated by another mechanism than eIF-4E phosphorylation. By using P19 cells overexpressing the EGF receptor, we show that the signal transduction pathway that leads to phosphorylation of eIF-4E is present in P19 cells; the EGF-induced change in phosphorylation of eIF-4E in these cells is likely to be regulated by a change in eIF-4E phosphatase activity. These results suggest that the onset of retinoic acid-induced differentiation is triggered by a signal transduction pathway which involves changes in protein synthesis, but not eIF-4E phosphorylation.