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. 1973 Jan;113(1):96–104. doi: 10.1128/jb.113.1.96-104.1973

Temperature-Sensitive Mutant of Escherichia coli K-12 with an Impaired d-Alanine: d-Alanine Ligase

E J J Lugtenberg a,1, Arna van Schijndel-van Dam a
PMCID: PMC251607  PMID: 4567144

Abstract

The temperature-sensitive Escherichia coli mutant strain ST-640 lyses at the restrictive temperature except when an osmotic stabilizer or a high concentration of d-alanine is present. The presence of dl-alanyl-dl-alanine does not prevent lysis. The rate of murein synthesis, followed in a wall medium, is decreased at both 30 and 42 C. d-Alanyl-d-alanine and uridine diphosphate-N-acetyl-muramyl (UDP-MurNAc)-pentapeptide are synthesized in decreased amounts, accompanied by accumulation of UDP-MurNAc-tripeptide at 42 C but not at 30 C. Uridine nucleotide precursors leak into the medium, especially out of the mutant cells. This leakage is prevented when NaCl is present. The d-alanine: d-alanine ligase (ADP) (EC 6.3.2.4) of the mutant strain, assayed in crude extracts, is temperature sensitive. The impaired ligase is relatively resistant to d-cycloserine and other inhibitors of the enzyme. Combined genetic and enzymatic results show that the low ligase activity is due to a mutation in the ddl gene, the structural gene for d-alanine: d-alanine ligase.

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Selected References

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

  1. Hendlin D., Stapley E. O., Jackson M., Wallick H., Miller A. K., Wolf F. J., Miller T. W., Chaiet L., Kahan F. M., Foltz E. L. Phosphonomycin, a new antibiotic produced by strains of streptomyces. Science. 1969 Oct 3;166(3901):122–123. doi: 10.1126/science.166.3901.122. [DOI] [PubMed] [Google Scholar]
  2. Lugtenberg E. J., De Haas-Menger L., Ruyters W. H. Murein synthesis and identification of cell wall precursors of temperature-sensitive lysis mutants of Escherichia coli. J Bacteriol. 1972 Jan;109(1):326–335. doi: 10.1128/jb.109.1.326-335.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Lugtenberg E. J. Studies on Escherichia coli enzymes involved in the synthesis of uridine diphosphate-N-acetyl-muramyl-pentapeptide. J Bacteriol. 1972 Apr;110(1):26–34. doi: 10.1128/jb.110.1.26-34.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Lugtenberg E. J., de Haan P. G. A simple method for following the fate of alanine-containing components in murein synthesis in Escherichia coli. Antonie Van Leeuwenhoek. 1971;37(4):537–552. doi: 10.1007/BF02218524. [DOI] [PubMed] [Google Scholar]
  5. Lugtenberg E. J., v Schijndel-van Dam A. Temperature-sensitive mutants of Escherichia coli K-12 with low activities of the L-alanine adding enzyme and the D-alanyl-D-alanine adding enzyme. J Bacteriol. 1972 Apr;110(1):35–40. doi: 10.1128/jb.110.1.35-40.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Lugtenberg E. J., v Schijndel-van Dam A. Temperature-sensitive mutants of Escherichia coli K-12 with low activity of the diaminopimelic acid adding enzyme. J Bacteriol. 1972 Apr;110(1):41–46. doi: 10.1128/jb.110.1.41-46.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Matsuzawa H., Matsuhashi M., Oka A., Sugino Y. Genetic and biochemical studies on cell wall peptidoglycan synthesis in Escherichia coli K-12. Biochem Biophys Res Commun. 1969 Aug 15;36(4):682–689. doi: 10.1016/0006-291x(69)90360-x. [DOI] [PubMed] [Google Scholar]
  8. Strominger J. L., Izaki K., Matsuhashi M., Tipper D. J. Peptidoglycan transpeptidase and D-alanine carboxypeptidase: penicillin-sensitive enzymatic reactions. Fed Proc. 1967 Jan-Feb;26(1):9–22. [PubMed] [Google Scholar]
  9. Taylor A. L. Current linkage map of Escherichia coli. Bacteriol Rev. 1970 Jun;34(2):155–175. doi: 10.1128/br.34.2.155-175.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Venkateswaran P. S., Wu H. C. Isolation and characterization of a phosphonomycin-resistant mutant of Escherichia coli K-12. J Bacteriol. 1972 Jun;110(3):935–944. doi: 10.1128/jb.110.3.935-944.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]

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