Effects of specific inhibitors on the coordination of the concentrations of ribulose-bisphosphate-carboxylase subunits and their corresponding mRNAs in the alga Chlorogonium

Abstract

Investigations were carried out on the effects of inhibitors of transcription and translation on the concentrations of the subunits of the plastid enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCase) and their corresponding mRNAs in the unicellular green alga Chlorogonium elongatum Dangeard. The light-induced increase of nuclear-coded small-subunit mRNA was strongly inhibited by α-amanitin while the increase of plastid-coded large-subunit mRNA was only weakly affected: Consequently, the mRNAs of the two subunits were present in very different proportions. Nevertheless, the light-induced increase of both subunits was strongly reduced by α-amanitin to the same degree, and hence the ratio of their concentrations was not affected compared with the untreated control cells. The effect of cycloheximide on the subunit mRNAs was similar to but weaker than that of α-amanitin. Again the increases in the subunit levels were strongly inhibited to the same degree. By contrast, rifampicin and chloramphenicol inhibited the light-induced increase of large-subunit mRNA more strongly than that of small-subunit mRNA, but the differences were less distinct than those caused by α-amanitin and cycloheximide. Again, the increase in both subunits was inhibited almost to the same extent. These results - especially those of the α-amanitin experiments - clearly show that the fine coordination of the RuBPCase subunits occurs posttranscriptionally at the level of translation and-or degradation. This conclusion was confirmed by pulse-chase experiments. Inhibition of the synthesis of the large subunits by chloramphenicol resulted - as also found by other authors-in a degradation of excess small subunits in the plastid. On the other hand, inhibition of the concentration of small subunits caused a proportionate reduction in the synthesis of large subunits, but no rapid degradation of large subunits could be detected. Therefore, the fine coordination of both subunits of RuBPCase is achieved by the degradation of an excess of small subunits, while the level of large subunits is adapted to the small subunit concentration, probably by adjustment of translation of the large-subunit mRNA. Furthermore, our experiments with α-amanitin and cycloheximide allow us to conclude that in the blue-light induction of large-subunit mRNA in the plastid the nucleocytoplasmic compartment is not directly involved.