Ribulose 1,5-Diphosphate Carboxylase Synthesis in Euglena: II. Effect of Inhibitors on Enzyme Synthesis during Regreening and Subsequent Transfer to Darkness.

Dark-grown Euglena gracilis Klebs strain Z Pringsheim cells, which have been partially regreened in the light, show a striking, continued synthesis of the chloroplast enzyme ribulose 1,5-diphosphate carboxylase on transfer back into darkness. This dark synthesis of the enzyme was completely prevented by the addition of 15 mug/ml of cycloheximide to the culture medium but was unaffected, for at least 8 hours, by the addition of 1 mg/ml of d-threo-chloramphenicol. The addition of either cycloheximide or d-threo-chloramphenicol to dark-grown cultures at the onset of illumination completely inhibited the light-induced synthesis of ribulose 1,5-diphosphate carboxylase. When cells which had been illuminated in the presence of d-threo-chloramphenicol, and hence were unable to synthesize ribulose 1,5-diphosphate carboxylase, were transferred to darkness in the absence of this inhibitor, synthesis of the carboxylase then occurred. Dark-grown cells which had been illuminated in the presence of cycloheximide failed to synthesize the enzyme when placed in the dark in the absence of cycloheximide. The addition of 5-fluorouracil to regreening cultures to prevent light-induced transcriptional steps completely blocked the synthesis of ribulose 1,5-diphosphate carboxylase.