Influence of growth conditions on RNA levels in relation to activity of core metabolic enzymes in the parasitic protists Trypanosoma brucei and Trichomonas vaginalis

Abstract

Levels of mRNAs encoding metabolic enzymes and their cellular activities were measured on continuous culture samples of the parasitic protists Trypanosoma brucei and Trichomonas vaginalis. The organisms were grown in chemostats at varying growth rates under glucose limitation or in the presence of excess glucose (EG), resulting in extensive adaptation of the cellular activities of glycolytic enzymes. rRNA and mRNA for beta-tubulin were monitored as controls. In Trypanosoma brucei levels of all RNAs showed a biphasic dependence on growth rate (= dilution rate D), with a sharp increase at higher D values. Cellular RNA levels of Trichomonas vaginalis rate-limited by glucose decreased slightly with increasing D. In EG-grown cells the opposite trend was observed. Equal levels for both carbon regimes were observed at intermediate D values. In both species the ratio between rRNA and mRNA encoding beta-tubulin was constant, independent of the carbon regime. mRNA encoding metabolic enzymes showed varying degrees of correlation with rRNA and beta-tubulin mRNA. In contrast, there was little to no correlation between mRNA levels and the activities of the enzymes they encode, even though only one of these is allosterically regulated. The data indicate that RNA levels in Trypanosoma brucei and Trichomonas vaginalis are determined by growth rate and in the latter species by the availability of the carbon and energy source. Rates of synthesis of metabolic enzymes are most likely regulated at the post-transcriptional level.