Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1990 Jun;172(6):3060–3065. doi: 10.1128/jb.172.6.3060-3065.1990

Acetohydroxy acid synthase activity from a mutation at ilvF in Escherichia coli K-12.

C Alexander-Caudle 1, L M Latinwo 1, J H Jackson 1
PMCID: PMC209108  PMID: 2188950

Abstract

Examination of the ilvF locus at 54 min on the Escherichia coli K-12 chromosome revealed that it is a cryptic gene for expression of a valine-resistant acetohydroxy acid synthase (acetolactate synthase; EC 4.1.3.18) distinct from previously reported isozymes. A spontaneous mutation, ilvF663, yielded IlvF+ enzyme activity that was multivalently repressed by all three branched-chain amino acids, was completely insensitive to feedback inhibition, was highly stable at elevated temperatures, and expressed optimal activity at 50 degrees C. The IlvF+ enzyme activity was expressed in strains in which isozyme II was inactive because of the ilvG frameshift in the wild-type strain K-12 and isozymes I and III were inactivated by point mutations or deletions. Tn5 insertional mutagenesis yielded two IlvF- mutants, with the insertion in ilvF663 in each case. These observations suggest that the ilvF663 locus may be a coding region for a unique acetohydroxy acid synthase activity.

Full text

PDF
3060

Selected References

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

  1. BEERS R. F., Jr, SIZER I. W. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem. 1952 Mar;195(1):133–140. [PubMed] [Google Scholar]
  2. 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]
  3. Blatt J. M., Pledger W. J., Umbarger H. E. Isoleucine and valine metabolism in Escherichia coli. XX. Multiple forms of acetohydroxy acid synthetase. Biochem Biophys Res Commun. 1972 Jul 25;48(2):444–450. doi: 10.1016/s0006-291x(72)80071-8. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Chang Y. Y., Cronan J. E., Jr Common ancestry of Escherichia coli pyruvate oxidase and the acetohydroxy acid synthases of the branched-chain amino acid biosynthetic pathway. J Bacteriol. 1988 Sep;170(9):3937–3945. doi: 10.1128/jb.170.9.3937-3945.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cocks G. T., Aguilar T., Lin E. C. Evolution of L-1, 2-propanediol catabolism in Escherichia coli by recruitment of enzymes for L-fucose and L-lactate metabolism. J Bacteriol. 1974 Apr;118(1):83–88. doi: 10.1128/jb.118.1.83-88.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dailey F. E., Cronan J. E., Jr Acetohydroxy acid synthase I, a required enzyme for isoleucine and valine biosynthesis in Escherichia coli K-12 during growth on acetate as the sole carbon source. J Bacteriol. 1986 Feb;165(2):453–460. doi: 10.1128/jb.165.2.453-460.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davis E. J., Blatt J. M., Henderson E. K., Whittaker J. J., Jackson J. H. Valine-sensitive acetohydroxy acid synthases in Escherichia coli K-12: unique regulation modulated by multiple genetic sites. Mol Gen Genet. 1977 Nov 18;156(3):239–249. doi: 10.1007/BF00267178. [DOI] [PubMed] [Google Scholar]
  9. De Felice M., Guardiola J., Esposito B., Iaccarino M. Structural genes for a newly recognized acetolactate synthase in Escherichia coli K-12. J Bacteriol. 1974 Dec;120(3):1068–1077. doi: 10.1128/jb.120.3.1068-1077.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Driver R. P., Lawther R. P. Physical analysis of deletion mutations in the ilvGEDA operon of Escherichia coli K-12. J Bacteriol. 1985 May;162(2):598–606. doi: 10.1128/jb.162.2.598-606.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hacking A. J., Aguilar J., Lin E. C. Evolution of propanediol utilization in Escherichia coli: mutant with improved substrate-scavenging power. J Bacteriol. 1978 Nov;136(2):522–530. doi: 10.1128/jb.136.2.522-530.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hacking A. J., Lin E. C. Disruption of the fucose pathway as a consequence of genetic adaptation to propanediol as a carbon source in Escherichia coli. J Bacteriol. 1976 Jun;126(3):1166–1172. doi: 10.1128/jb.126.3.1166-1172.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hall B. G., Yokoyama S., Calhoun D. H. Role of cryptic genes in microbial evolution. Mol Biol Evol. 1983 Dec;1(1):109–124. doi: 10.1093/oxfordjournals.molbev.a040300. [DOI] [PubMed] [Google Scholar]
  14. Jackson J. H. Rapid assay of acetolactate synthase in permeabilized bacteria. Methods Enzymol. 1988;166:230–233. doi: 10.1016/s0076-6879(88)66030-7. [DOI] [PubMed] [Google Scholar]
  15. Lawther R. P., Calhoun D. H., Adams C. W., Hauser C. A., Gray J., Hatfield G. W. Molecular basis of valine resistance in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1981 Feb;78(2):922–925. doi: 10.1073/pnas.78.2.922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Newman T. C., Levinthal M. A new map location for the ilvB locus of Escherichia coli. Genetics. 1980 Sep;96(1):59–77. doi: 10.1093/genetics/96.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. O'Neill J. P., Freundlich M. Two forms of biosynthetic acetohydroxy acid synthetase in Salmonella typhimurium. Biochem Biophys Res Commun. 1972 Jul 25;48(2):437–443. doi: 10.1016/s0006-291x(72)80070-6. [DOI] [PubMed] [Google Scholar]
  18. Pledger W. J., Umbarger H. E. Isoleucine and valine metabolism in Escherichia coli. XXI. Mutations affecting derepression and valine resistance. J Bacteriol. 1973 Apr;114(1):183–194. doi: 10.1128/jb.114.1.183-194.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Riley M., Anilionis A. Evolution of the bacterial genome. Annu Rev Microbiol. 1978;32:519–560. doi: 10.1146/annurev.mi.32.100178.002511. [DOI] [PubMed] [Google Scholar]
  20. Robinson C. L., Jackson J. H. New acetohydroxy acid synthase activity from mutational activation of a cryptic gene in Escherichia coli K-12. Mol Gen Genet. 1982;186(2):240–246. doi: 10.1007/BF00331856. [DOI] [PubMed] [Google Scholar]
  21. Rosner J. L. Formation, induction, and curing of bacteriophage P1 lysogens. Virology. 1972 Jun;48(3):679–689. doi: 10.1016/0042-6822(72)90152-3. [DOI] [PubMed] [Google Scholar]

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

RESOURCES