Goldfine, HowardDaldal, Fevzi2022-09-152022-09-152007-12Aygün, S. S. vd. (2007). "Rhodobacter capsulatus O1sA is a bifunctional enyzme active in both ornithine lipid and phosphatidic acid biosynthesis". Journal of Bacteriology, 189(23), 8564-8574.0021-9193https://doi.org/10.1128/JB.01121-07https://journals.asm.org/doi/10.1128/JB.01121-07http://hdl.handle.net/11452/28753The Rhodobacter capsulatus genome contains three genes (olsA [plsC138], plsC316, and plsC3498) that are annotated as lysophosphatidic acid (1-acyl-sn-glycerol-3-phosphate) acyltransferase (AGPAT). Of these genes, olsA was previously shown to be an O-acyltransferase in the second step of ornithine lipid biosynthesis, which is important for optimal steady-state levels of c-type cytochromes (S. Aygun-Sunar, S. Mandaci, H.-G. Koch, I. V. J. Murray, H. Goldfine, and F. Daldal. Mol. Microbiol. 61:418-435, 2006). The roles of the remaining plsC316 and plsC3498 genes remained unknown. In this work, these genes were cloned, and chromosomal insertion-deletion mutations inactivating them were obtained to define their function. Characterization of these mutants indicated that, unlike the Escherichia coli plsC, neither plsC316 nor plsC3498 was essential in R. capsulatus. In contrast, no plsC316 olsA double mutant could be isolated, indicating that an intact copy of either olsA or plsC316 was required for R. capsulatus growth under the conditions tested. Compared to OlsA null mutants, PlsC316 null mutants contained ornithine lipid and had no c-type cytochrome-related phenotype. However, they exhibited slight growth impairment and highly altered total fatty acid and phospholipid profiles. Heterologous expression in an E. coli plsC(Ts) mutant of either R. capsulatus plsC316 or olsA gene products supported growth at a nonpermissive temperature, exhibited AGPAT activity in vitro, and restored phosphatidic acid biosynthesis. The more vigorous AGPAT activity displayed by PlsC316 suggested that plsC316 encodes the main AGPAT required for glycerophospholipid synthesis in R. capsulatus, while olsA acts as an alternative AGPAT that is specific for ornithine lipid synthesis. This study therefore revealed for the first time that some OlsA enzymes, like the enzyme of R. capsulatus, are bifunctional and involved in both membrane ornithine lipid and glycerophospholipid biosynthesis.eninfo:eu-repo/semantics/openAccessEscherichia coliAcyl carrier proteinRhodobacter capsulatusEscherichia-coli1-acyl-sn-glycerol-3-phosphate acyltransferasePhospholipid-synthesisGlycerophospholipid biosynthesisRhodopseudomonas-capsulataNeisseria-meningitidisRhizobium-melilotiCdna cloningGeneGlycerol-3-phosphate O-acyltransferase1-Acylglycerol-3-Phosphate O-AcyltransferaseAmino acid sequenceBacterial proteinsCytochromes cGene expression regulation, bacterialGlycerophospholipidsMutationMolecular sequence dataLipidsOpen reading framesOrnithinePhosphatidic acidsRhodobacter capsulatusArticle0002514841000182-s2.0-36749036019856485741892317921310MicrobiologyOrnithine Containing Aminolipid; Glucuronosyl Diacylglycerol; Membrane LipidsBiosynthesisLysophosphatidic acidAcyltransferaseGene productOrnithinePhosphatidic acidArticleBacterial geneBacterial growthBacterial strainEnzyme activationPriority journalControlled studyGene insertionEnzyme activityGene deletionGene functionMolecular cloningRhodobacter capsulatusIn vitro studyNonhumanOlsa genePlsc316 genePlsc3498 gene