Abstract
Unsaturated fatty acid auxotrophs of Escherichia coli are able to use only unsaturated fatty acids of the cis configuration as the required growth supplement. A mutation in the fatA gene allows such auxotrophs to utilize unsaturated fatty acids with a trans double bond as well as fatty acids having a cis double bond. The fatA gene was mapped to min 69 near argG, and the allele studied (fatA1) was found to be dominant over the wild-type gene. fatA1 mutant strains grew at similar rates when supplemented with elaidate (trans-9-octadecenoate) or oleate (cis-9-octadecenoate). The fat+ strain, however, lysed when supplemented with the trans fatty acid. Physiological characterization of the fatA mutant strain was undertaken. The mutation appeared not to be involved with long-chain fatty acid transport. Introduction of lesions in known fatty acid transport genes abolished trans fatty acid utilization in the fatA mutant strain. Also, growth characteristics of the fat+ and the fatA1 mutant strains on elaidate as the sole carbon source were identical, which indicated comparables rate of fatty acid accumulation. The mutation appeared to be involved with recognition of the trans configuration after uptake into the cell. The levels of trans fatty acid incorporation into the phospholipids of the fat+ and the fatA strains differed considerably, with the mutant incorporating much higher levels. No significant accumulation of elaidate into nonphospholipid cellular components was observed. The fatA mutation did not appear to be involved with the cellular metabolic state, as cyclic AMP had no effect on the ability of the strains to utilize trans fatty acids.
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- BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
- Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chang Y. Y., Cronan J. E., Jr Genetic and biochemical analyses of Escherichia coli strains having a mutation in the structural gene (poxB) for pyruvate oxidase. J Bacteriol. 1983 May;154(2):756–762. doi: 10.1128/jb.154.2.756-762.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clark D. Regulation of fatty acid degradation in Escherichia coli: analysis by operon fusion. J Bacteriol. 1981 Nov;148(2):521–526. doi: 10.1128/jb.148.2.521-526.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cronan J. E., Jr, Gelmann E. P. Physical properties of membrane lipids: biological relevance and regulation. Bacteriol Rev. 1975 Sep;39(3):232–256. doi: 10.1128/br.39.3.232-256.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cronan J. E., Jr Regulation of the fatty acid composition of the membrane phospholipids of Escherichia coli. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3758–3762. doi: 10.1073/pnas.71.9.3758. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cronan J. E., Jr, Silbert D. F., Wulff D. L. Mapping of the fabA locus for unsaturated fatty acid biosynthesis in Escherichia coli. J Bacteriol. 1972 Oct;112(1):206–211. doi: 10.1128/jb.112.1.206-211.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cronan J. E., Jr Thermal regulation of the membrane lipid composition of Escherichia coli. Evidence for the direct control of fatty acid synthesis. J Biol Chem. 1975 Sep 10;250(17):7074–7077. [PubMed] [Google Scholar]
- Esfahani M., Barnes E. M., Jr, Wakil S. J. Control of fatty acid composition in phospholipids of Escherichia coli: response to fatty acid supplements in a fatty acid auxotroph. Proc Natl Acad Sci U S A. 1969 Nov;64(3):1057–1064. doi: 10.1073/pnas.64.3.1057. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Esfahani M., Ioneda T., Wakil S. J. Studies on the control of fatty acid metabolism. 3. Incorporation of fatty acids into phospholipids and regulation of fatty acid synthetase of Escherichia coli. J Biol Chem. 1971 Jan 10;246(1):50–56. [PubMed] [Google Scholar]
- Fox C. F., Law J. H., Tsukagoshi N., Wilson G. A density label for membranes. Proc Natl Acad Sci U S A. 1970 Oct;67(2):598–605. doi: 10.1073/pnas.67.2.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gally H. U., Pluschke G., Overath P., Seelig J. Structure of Escherichia coli membranes. Fatty acyl chain order parameters of inner and outer membranes and derived liposomes. Biochemistry. 1980 Apr 15;19(8):1638–1643. doi: 10.1021/bi00549a018. [DOI] [PubMed] [Google Scholar]
- Garwin J. L., Klages A. L., Cronan J. E., Jr Structural, enzymatic, and genetic studies of beta-ketoacyl-acyl carrier protein synthases I and II of Escherichia coli. J Biol Chem. 1980 Dec 25;255(24):11949–11956. [PubMed] [Google Scholar]
- Grabau C., Cronan J. E., Jr In vivo function of Escherichia coli pyruvate oxidase specifically requires a functional lipid binding site. Biochemistry. 1986 Jul 1;25(13):3748–3751. doi: 10.1021/bi00361a003. [DOI] [PubMed] [Google Scholar]
- Haest C. W., de Gier J., van Es G. A., Verkleij A. J., van Deenen L. L. Fragility of the permeability barrier of Escherichia coli. Biochim Biophys Acta. 1972 Oct 23;288(1):43–53. doi: 10.1016/0005-2736(72)90221-0. [DOI] [PubMed] [Google Scholar]
- Kleckner N., Roth J., Botstein D. Genetic engineering in vivo using translocatable drug-resistance elements. New methods in bacterial genetics. J Mol Biol. 1977 Oct 15;116(1):125–159. doi: 10.1016/0022-2836(77)90123-1. [DOI] [PubMed] [Google Scholar]
- Klein K., Steinberg R., Fiethen B., Overath P. Fatty acid degradation in Escherichia coli. An inducible system for the uptake of fatty acids and further characterization of old mutants. Eur J Biochem. 1971 Apr;19(3):442–450. doi: 10.1111/j.1432-1033.1971.tb01334.x. [DOI] [PubMed] [Google Scholar]
- Maloy S. R., Ginsburgh C. L., Simons R. W., Nunn W. D. Transport of long and medium chain fatty acids by Escherichia coli K12. J Biol Chem. 1981 Apr 25;256(8):3735–3742. [PubMed] [Google Scholar]
- Narasimhan M. L., Lampi J. L., Cronan J. E., Jr Genetic and biochemical characterization of an Escherichia coli K-12 mutant deficient in acyl-coenzyme A thioesterase II. J Bacteriol. 1986 Mar;165(3):911–917. doi: 10.1128/jb.165.3.911-917.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nunn W. D. A molecular view of fatty acid catabolism in Escherichia coli. Microbiol Rev. 1986 Jun;50(2):179–192. doi: 10.1128/mr.50.2.179-192.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nunn W. D., Colburn R. W., Black P. N. Transport of long-chain fatty acids in Escherichia coli. Evidence for role of fadL gene product as long-chain fatty acid receptor. J Biol Chem. 1986 Jan 5;261(1):167–171. [PubMed] [Google Scholar]
- Nunn W. D., Simons R. W. Transport of long-chain fatty acids by Escherichia coli: mapping and characterization of mutants in the fadL gene. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3377–3381. doi: 10.1073/pnas.75.7.3377. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Overath P., Brenner M., Gulik-Krzywicki T., Shechter E., Letellier L. Lipid phase transitions in cytoplasmic and outer membranes of Escherichia coli. Biochim Biophys Acta. 1975 May 6;389(2):358–369. doi: 10.1016/0005-2736(75)90328-4. [DOI] [PubMed] [Google Scholar]
- Overath P., Hill F. F., Lamnek-Hirsch I. Biogenesis of E. coli membrane: evidence for randomization of lipid phase. Nat New Biol. 1971 Dec 29;234(52):264–267. doi: 10.1038/newbio234264a0. [DOI] [PubMed] [Google Scholar]
- Overath P., Pauli G., Schairer H. U. Fatty acid degradation in Escherichia coli. An inducible acyl-CoA synthetase, the mapping of old-mutations, and the isolation of regulatory mutants. Eur J Biochem. 1969 Feb;7(4):559–574. [PubMed] [Google Scholar]
- Overath P., Schairer H. U., Stoffel W. Correlation of in vivo and in vitro phase transitions of membrane lipids in Escherichia coli. Proc Natl Acad Sci U S A. 1970 Oct;67(2):606–612. doi: 10.1073/pnas.67.2.606. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Polacco M. L., Cronan J. E., Jr Mechanism of the apparent regulation of Escherichia coli unsaturated fatty acid synthesis by exogenous oleic acid. J Biol Chem. 1977 Aug 10;252(15):5488–5490. [PubMed] [Google Scholar]
- Simons R. W., Egan P. A., Chute H. T., Nunn W. D. Regulation of fatty acid degradation in Escherichia coli: isolation and characterization of strains bearing insertion and temperature-sensitive mutations in gene fadR. J Bacteriol. 1980 May;142(2):621–632. doi: 10.1128/jb.142.2.621-632.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsao Y. K., Lands W. E. Cell growth with trans fatty acids is affected by adenosine 3',5'-monophosphate and membrane fluidity. Science. 1980 Feb 15;207(4432):777–779. doi: 10.1126/science.6243419. [DOI] [PubMed] [Google Scholar]
- Ulrich A. K., de Mendoza D., Garwin J. L., Cronan J. E., Jr Genetic and biochemical analyses of Escherichia coli mutants altered in the temperature-dependent regulation of membrane lipid composition. J Bacteriol. 1983 Apr;154(1):221–230. doi: 10.1128/jb.154.1.221-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vallari D. S., Rock C. O. Isolation and characterization of Escherichia coli pantothenate permease (panF) mutants. J Bacteriol. 1985 Oct;164(1):136–142. doi: 10.1128/jb.164.1.136-142.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilson G., Fox C. F. Biogenesis of microbial transport systems: evidnce for coupled incorporation of newly synthesized lipids and proteins into membrane. J Mol Biol. 1971 Jan 14;55(1):49–60. doi: 10.1016/0022-2836(71)90280-4. [DOI] [PubMed] [Google Scholar]
- Wilson G., Rose S. P., Fox C. F. The effect of membrane lipid unsaturation on glycoside transport. Biochem Biophys Res Commun. 1970 Feb 20;38(4):617–623. doi: 10.1016/0006-291x(70)90625-x. [DOI] [PubMed] [Google Scholar]

