Mechanisms of regulation of the biosynthesis of membrane-derived oligosaccharides in Escherichia coli.

Abstract

The periplasmic glucans of Gram-negative bacteria, including the membrane-derived oligosaccharides (MDO) of Escherichia coli and the cyclic glucans of the Rhizobiaceae, have important but poorly understood functions in osmotic adaptation and, in the case of the Rhizobiaceae, in the complex cell-signaling of these bacteria with specific plant hosts. Experiments on the mechanisms of osmotic regulation of the biosynthesis of MDO in E. coli reported here support a model in which osmotic regulation occurs principally at the level of modulation of enzyme activity rather than at the level of gene expression. 1) Activity of the membrane-bound glucosyltransferase thought to catalyze the first and rate-making step in the biosynthesis of MDO is not altered by the osmolarity of the medium in which cells are grown. 2) Upon dilution of cells growing at high osmolarity into a medium of low osmolarity, the increased synthesis of MDO begins at maximum rate without detectable lag. 3) The activity of the membrane glucosyltransferase in vitro is strongly inhibited by high levels of salts, consistent with the view that synthesis in vivo is regulated chiefly by this mechanism, rather than by regulation of the synthesis of biosynthetic enzymes. We also find that the biosynthesis of MDO is regulated not only osmotically but also by strong feedback inhibition in response to the levels of MDO in the periplasm.