Regulation of synthesis of citrate synthase in regreening Euglena gracilis

Abstract

Exposure of dark‐grown Euglena to white or red light, but not blue light, produced a twofold increase in the specific activity of citrate synthase. A 400‐fold purification of mitochondrial citrate synthase (subunit Mr= 44000) was achieved from cells of Euglena gracilis by affinity chromatography on ATP‐activated agarose. Antisera, raised against the homogeneously pure enzyme, were used to demonstrate that the increase in citrate synthase activity on exposure of dark‐grown cells to light resulted from an increase in citrate synthase protein. Anti‐(citrate synthase was used to detect precursor citrate synthase resulting from the translation of total polyadenylated RNA from Euglena in a cell‐free rabbit reticulocyte lysate system. Citrate synthase mRNA was found to be present in cells at all stages of regreening. However, extraction and translation of polyadenylated RNA from free polysomes isolated from dark‐grown and regreening cells demonstrated that appreciable translation of citrate synthase mRNA was only occurring in regreening cells. Copyright © 1984, Wiley Blackwell. All rights reserved