Ethanol-induced reduction of the amount of stimulatory GTP-binding regulatory protein, Gs, in wild-type S49 murine lymphoma cells: effect of cycloheximide and puromycin.

The activity and the amount of Gs in wild-type S49 murine lymphoma cells decreased by 16-18% and by 29%, respectively, after 24 h treatment of the cells with 50 mM ethanol, through a transient increase of the Gs activity with no alteration in its content after 4 and 8 h treatment. No decrease of the Gs activity was noted when membranes from S49 cells were treated with 50-500 mM ethanol. To study the role of protein metabolism in the ethanol-induced reduction in the amount of Gs, S49 cells were treated with ethanol for 24 h in the presence of inhibitor of protein synthesis. Cycloheximide at concentrations of 0.05-0.15 micrograms/ml and puromycin at 0.15-0.60 micrograms/ml caused a dose-dependent inhibition of cell division in S49 cells to a similar degree. The ethanol-induced reduction of the activity and the amount of Gs was completely abolished by those concentrations of cycloheximide, but not by puromycin. Cycloheximide itself caused no alteration in the Gs activity of S49 cell membranes. These results indicate that the process of the ethanol-induced event of Gs involves cycloheximide-sensitive protein synthesis.