Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Sep;178(18):5382–5387. doi: 10.1128/jb.178.18.5382-5387.1996

Increased unsaturated fatty acid production associated with a suppressor of the fabA6(Ts) mutation in Escherichia coli.

C O Rock 1, J T Tsay 1, R Heath 1, S Jackowski 1
PMCID: PMC178354  PMID: 8808925

Abstract

Plasmids that corrected the temperature-sensitive unsaturated fatty acid auxotrophy of strain M6 [fabA6 (Ts)] were isolated from an Escherichia coli genomic library. Subcloning and physical mapping localized the new gene (called sfa for suppressor of fabA) at 1,070 kb on the E. coli chromosome. DNA sequencing revealed the presence of a 227-bp open reading frame which directed the synthesis of a peptide of approximately 8 kDa, which correlated with the correction of the fabA6(Ts) phenotype. However, the sfa gene was an allele-specific suppressor since plasmids harboring the sfa gene corrected the growth phenotype of fabA6(Ts) mutants but did not correct the growth of fabA2(Ts) or fabB15(Ts) unsaturated fatty acid auxotrophs. Overexpression of the sfa gene in fabA6(Ts) mutants restored unsaturated fatty acid content at 42 degrees C, and overexpression in wild-type cells resulted in a substantial increase in the unsaturated fatty acid content of the membrane. Thus, the suppression of the fabA6(Ts) mutation by sfa was attributed to its ability to increase the biosynthesis of unsaturated fatty acids.

Full Text

The Full Text of this article is available as a PDF (332.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aiba H., Adhya S., de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed] [Google Scholar]
  2. 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]
  3. Clark D. P., DeMendoza D., Polacco M. L., Cronan J. E., Jr Beta-hydroxydecanoyl thio ester dehydrase does not catalyze a rate-limiting step in Escherichia coli unsaturated fatty acid synthesis. Biochemistry. 1983 Dec 6;22(25):5897–5902. doi: 10.1021/bi00294a032. [DOI] [PubMed] [Google Scholar]
  4. Clark D., Cronan J. E., Jr Further mapping of several membrane lipid biosynthetic genes (fabC, fabB, gpsA, plsB) of Escherichia coli. J Bacteriol. 1977 Nov;132(2):549–554. doi: 10.1128/jb.132.2.549-554.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cooper C. L., Jackowski S., Rock C. O. Fatty acid metabolism in sn-glycerol-3-phosphate acyltransferase (plsB) mutants. J Bacteriol. 1987 Feb;169(2):605–611. doi: 10.1128/jb.169.2.605-611.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cronan J. E., Jr, Birge C. H., Vagelos P. R. Evidence for two genes specifically involved in unsaturated fatty acid biosynthesis in Escherichia coli. J Bacteriol. 1969 Nov;100(2):601–604. doi: 10.1128/jb.100.2.601-604.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Cronan J. E., Jr, Li W. B., Coleman R., Narasimhan M., de Mendoza D., Schwab J. M. Derived amino acid sequence and identification of active site residues of Escherichia coli beta-hydroxydecanoyl thioester dehydrase. J Biol Chem. 1988 Apr 5;263(10):4641–4646. [PubMed] [Google Scholar]
  9. 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]
  10. DiRusso C. C., Nunn W. D. Cloning and characterization of a gene (fadR) involved in regulation of fatty acid metabolism in Escherichia coli. J Bacteriol. 1985 Feb;161(2):583–588. doi: 10.1128/jb.161.2.583-588.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Heath R. J., Rock C. O. Enoyl-acyl carrier protein reductase (fabI) plays a determinant role in completing cycles of fatty acid elongation in Escherichia coli. J Biol Chem. 1995 Nov 3;270(44):26538–26542. doi: 10.1074/jbc.270.44.26538. [DOI] [PubMed] [Google Scholar]
  12. Henry M. F., Cronan J. E., Jr A new mechanism of transcriptional regulation: release of an activator triggered by small molecule binding. Cell. 1992 Aug 21;70(4):671–679. doi: 10.1016/0092-8674(92)90435-f. [DOI] [PubMed] [Google Scholar]
  13. Henry M. F., Cronan J. E., Jr Escherichia coli transcription factor that both activates fatty acid synthesis and represses fatty acid degradation. J Mol Biol. 1991 Dec 20;222(4):843–849. doi: 10.1016/0022-2836(91)90574-p. [DOI] [PubMed] [Google Scholar]
  14. Jenkins L. S., Nunn W. D. Genetic and molecular characterization of the genes involved in short-chain fatty acid degradation in Escherichia coli: the ato system. J Bacteriol. 1987 Jan;169(1):42–52. doi: 10.1128/jb.169.1.42-52.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kauppinen S., Siggaard-Andersen M., von Wettstein-Knowles P. beta-Ketoacyl-ACP synthase I of Escherichia coli: nucleotide sequence of the fabB gene and identification of the cerulenin binding residue. Carlsberg Res Commun. 1988;53(6):357–370. doi: 10.1007/BF02983311. [DOI] [PubMed] [Google Scholar]
  16. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  17. Kotani H., Kawamura A., Takahashi A., Nakatsuji M., Hiraoka N., Nakajima K., Takanami M. Site-specific dissection of E. coli chromosome by lambda terminase. Nucleic Acids Res. 1992 Jul 11;20(13):3357–3360. doi: 10.1093/nar/20.13.3357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Magnuson K., Jackowski S., Rock C. O., Cronan J. E., Jr Regulation of fatty acid biosynthesis in Escherichia coli. Microbiol Rev. 1993 Sep;57(3):522–542. doi: 10.1128/mr.57.3.522-542.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rosenfeld I. S., D'Agnolo G., Vagelos P. R. Synthesis of unsaturated fatty acids and the lesion in fab B mutants. J Biol Chem. 1973 Apr 10;248(7):2452–2460. [PubMed] [Google Scholar]
  20. Silbert D. F., Vagelos P. R. Fatty acid mutant of E. coli lacking a beta-hydroxydecanoyl thioester dehydrase. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1579–1586. doi: 10.1073/pnas.58.4.1579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. de Mendoza D., Klages Ulrich A., Cronan J. E., Jr Thermal regulation of membrane fluidity in Escherichia coli. Effects of overproduction of beta-ketoacyl-acyl carrier protein synthase I. J Biol Chem. 1983 Feb 25;258(4):2098–2101. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES