Neutral lipid accumulation in the membranes of Escherichia coli mutants lacking diglyceride kinase

Abstract

We have developed a rapid autoradiographic screening assay for detecting diglyceride kinase in colonies of Escherichia coli and have isolated four strains lacking this enzyme. The gene (designated dgk) which is altered in these mutants is cotransduceable with the malB locus, near minute 90 on the chromosome. The membranes of strain RZ60 (which carries the dgk-6 lesion) contain substantial amounts of 1,2-diglyceride, representing approximately 8% of the total lipid. In contrast, wild type cells of E. coli (dgk+) only contain about 0.5% 1,2-diglyceride. The phospholipid composition of these mutants is not dramatically altered, and they are not temperature sensitive for growth. However, strains bearing the dgk-6 mutation do not grow well on nutrient media of low osmolarity. This can be corrected by the inclusion of 1% NaCl or 0.5 M sucrose. These results suggest that 1,2-diglyceride is the true substrate for the kinase in vivo and that the kinase functions as a minor route for phosphatidic acid synthesis. Genetic modification of the diglyceride content of the E. coli membrane has not been reported previously.