Genetic and biochemical analyses of Escherichia coli mutants altered in the temperature-dependent regulation of membrane lipid composition.
We have previously studied two mutants of Escherichia coli altered in the regulation of membrane lipid composition by temperature. One class (represented by the fabFl allele) fails to regulate upon temperature shift and is defective in cis-vaccenic acid synthesis owing to the lack of the fatty acid elongation enzyme beta-ketoacyl-acyl carrier protein synthase II(EC 2.3.1.41). A second class of mutant, given the phenotypic designation Vtr, overproduces cis-vaccenic acid at all temperatures and hence is altered in temperature regulation. In this paper we report evidence for the following conclusions. (i) The Vtr and fabFl mutations show very tight genetic linkage. (ii) The Vtr lesion is allelic to the fabFl mutation since the presence of the fabFl mutation in merodiploid strains carrying the Vtr or fabF(+) alleles results in fatty acid compositions intermediate between those of the two monoploid strains. Merodiploids carrying both the fabF(+) and Vtr alleles likewise show an intermediate composition. These results indicate intra-allelic complementation. (iii) The two E. coli proteins recently discovered by Rock (J. Bacteriol. 152:1298-1300, 1982) that form mixed disulfide cross-links to acyl carrier protein are directly demonstrated to be beta-ketoacyl-acyl carrier protein synthases I and II. (iv) The fabFl strains produce a synthase II band of altered electrophoretic mobility, indicating that the fabF locus is the structural gene for synthase II. (v) The synthase II of Vtr strains is abnormally sensitive to cerulenin, an antibiotic that specifically inhibits synthases I and II. This increased sensitivity is readily demonstrated in vivo, but in vitro we failed to detect an increased sensitivity of the Vtr synthase II to cerulenin, nor have we detected any other kinetic or structural alteration in the enzyme. We interpret these results in terms of specific interactions of synthase II with other cellular components which occur in vivo but are not duplicated in vitro.
D015099:3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / Q000235:genetics; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D008563:Membrane Lipids / Q000235:genetics*; D009154:Mutation; D013696:Temperature
D008563:Membrane Lipids; D015099:3-Oxoacyl-(Acyl-Carrier-Protein) Synthase