Phosphatidic acid accumulation in the membranes of Escherichia coli mutants defective in CDP-diglyceride synthetase.
CTP-phosphatidic acid cytidylyltransferase (CDP-diglyceride synthetase) is a key enzyme in the biogenesis of membrane phospholipids in Escherichia coli. Using a modification of a previously described autoradiographic screening procedure (Raetz, C. R. H. (1975) Proc. Natl. Acad. Sci. U.S.A. 72, 2274-2278), we have isolated six mutant strains in which the specific activity of the synthetase is 1 to 10% that of the wild type, as judged by in vitro assays. The synthesis of dCDP-diglyceride, as well as CDP-diglyceride, is defective in these organisms. The mutations responsible for the enzyme defects (designated cds) all map in the same location near minute 4 on the chromosome. Although none of the mutants obtained are temperature-sensitive for growth, all of them exhibit significantly elevated levels of phosphatidic acid in vivo. The highest increase is observed in the mutant GL60, in which phosphatidic acid constitutes about 5% of the membrane lipid, in contrast to 0.2% in typical wild type strains. The accumulation of phosphatidic acid occurs primarily at the expense of phosphatidylglycerol and cardiolipin, but the total lipid-to-protein ratio of GL60 is nearly normal. In vivo labeling of GL60 with 32Pi suggests that the increased phosphatidic acid pool is the result of a partial metabolic block early in the phospholipid pathway, but that most of this expanded pool is nonetheless available for de novo synthesis.
D002462:Cell Membrane / Q000378:metabolism; D003567:Cytidine Diphosphate Diglycerides; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005838:Genotype; D008563:Membrane Lipids / Q000378:metabolism*; D009154:Mutation; D009713:Nucleotidyltransferases / Q000235:genetics*; D010712:Phosphatidic Acids / Q000378:metabolism*; D013045:Species Specificity; D014161:Transduction, Genetic
D003567:Cytidine Diphosphate Diglycerides; D008563:Membrane Lipids; D010712:Phosphatidic Acids; D009713:Nucleotidyltransferases; C022348:phosphatidate cytidylyltransferase