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. 1983 Mar;153(3):1439–1450. doi: 10.1128/jb.153.3.1439-1450.1983

Two alanine racemase genes in Salmonella typhimurium that differ in structure and function.

S A Wasserman, C T Walsh, D Botstein
PMCID: PMC221795  PMID: 6298185

Abstract

Mutations were isolated in a previously undescribed Salmonella typhimurium gene encoding an alanine racemase essential for utilization of L-alanine as a source of carbon, energy, and nitrogen. This new locus, designated dadB, lies within one kilobase of the D-alanine dehydrogenase locus (dadA), which is also required for alanine catabolism. The dadA and dadB genes are coregulated. Mutants (including insertions) lacking the dadB alanine racemase do not require D-alanine for growth unless a mutation is introduced at a second locus, designated dal. Two genes specifying alanine racemase activity were cloned from S. typhimurium. The two cloned DNA sequences do not cross-hybridize with each other; one was shown to contain the dadB gene.

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  1. Atherton F. R., Hall M. J., Hassall C. H., Lambert R. W., Lloyd W. J., Ringrose P. S. Phosphonopeptides as antibacterial agents: mechanism of action of alaphosphin. Antimicrob Agents Chemother. 1979 May;15(5):696–705. doi: 10.1128/aac.15.5.696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berberich R., Kaback M., Freese E. D-amino acids as inducers of L-alanine dehydrogenase in Bacillus subtilis. J Biol Chem. 1968 Mar 10;243(5):1006–1011. [PubMed] [Google Scholar]
  3. Blattner F. R., Williams B. G., Blechl A. E., Denniston-Thompson K., Faber H. E., Furlong L., Grunwald D. J., Kiefer D. O., Moore D. D., Schumm J. W. Charon phages: safer derivatives of bacteriophage lambda for DNA cloning. Science. 1977 Apr 8;196(4286):161–169. doi: 10.1126/science.847462. [DOI] [PubMed] [Google Scholar]
  4. Bochner B. R., Savageau M. A. Generalized indicator plate for genetic, metabolic, and taxonomic studies with microorganisms. Appl Environ Microbiol. 1977 Feb;33(2):434–444. doi: 10.1128/aem.33.2.434-444.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bukhari A. I., Taylor A. L. Mutants of Escherichia coli with a growth requirement for either lysine or pyridoxine. J Bacteriol. 1971 Mar;105(3):988–998. doi: 10.1128/jb.105.3.988-998.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. FREESE E., PARK S. W., CASHEL M. THE DEVELOPMENTAL SIGNIFICANCE OF ALANINE DEHYDROGENASE IN BACILLUS SUBTILIS. Proc Natl Acad Sci U S A. 1964 Jun;51:1164–1172. doi: 10.1073/pnas.51.6.1164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Foster T. J., Davis M. A., Roberts D. E., Takeshita K., Kleckner N. Genetic organization of transposon Tn10. Cell. 1981 Jan;23(1):201–213. doi: 10.1016/0092-8674(81)90285-3. [DOI] [PubMed] [Google Scholar]
  8. Franklin F. C., Venables W. A. Biochemical, genetic, and regulatory studies of alanine catabolism in Escherichia coli K12. Mol Gen Genet. 1976 Dec 8;149(2):229–237. doi: 10.1007/BF00332894. [DOI] [PubMed] [Google Scholar]
  9. Hilliker S., Botstein D. An early regulatory gene of Salmonella phage P22 analogous to gene N of coliphage lambda. Virology. 1975 Dec;68(2):510–524. doi: 10.1016/0042-6822(75)90291-3. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Kollonitsch J., Barash L., Kahan F. M., Kropp H. Letter: New antibacterial agent via photofluorination of a bacterial cell wall constituent. Nature. 1973 Jun 8;243(5406):346–347. doi: 10.1038/243346a0. [DOI] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Lambert M. P., Neuhaus F. C. Factors affecting the level of alanine racemase in Escherichia coli. J Bacteriol. 1972 Mar;109(3):1156–1161. doi: 10.1128/jb.109.3.1156-1161.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lambert M. P., Neuhaus F. C. Mechanism of D-cycloserine action: alanine racemase from Escherichia coli W. J Bacteriol. 1972 Jun;110(3):978–987. doi: 10.1128/jb.110.3.978-987.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lugtenberg E. J., v Schijndel-van Dam A. Temperature-sensitive mutant of Escherichia coli K-12 with an impaired D-alanine:D-alanine ligase. J Bacteriol. 1973 Jan;113(1):96–104. doi: 10.1128/jb.113.1.96-104.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Manning J. M., Merrifield N. E., Jones W. M., Gotschlich E. C. Inhibition of bacterial growth by beta-chloro-D-alanine. Proc Natl Acad Sci U S A. 1974 Feb;71(2):417–421. doi: 10.1073/pnas.71.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Miyakawa T., Matsuzawa H., Matsuhashi M., Sugino Y. Cell wall peptidoglycan mutants of Escherichia coli K-12: existence of two clusters of genes, mra and mrb, for cell wall peptidoglycan biosynthesis. J Bacteriol. 1972 Nov;112(2):950–958. doi: 10.1128/jb.112.2.950-958.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mojica T. Transduction by phage P1CM clr-100 in Salmonella typhimurium. Mol Gen Genet. 1975;138(2):113–126. doi: 10.1007/BF02428116. [DOI] [PubMed] [Google Scholar]
  19. Murray N. E., Brammar W. J., Murray K. Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet. 1977 Jan 7;150(1):53–61. doi: 10.1007/BF02425325. [DOI] [PubMed] [Google Scholar]
  20. Neuhaus F. C., Hammes W. P. Inhibition of cell wall biosynthesis by analogues and alanine. Pharmacol Ther. 1981;14(3):265–319. doi: 10.1016/0163-7258(81)90030-9. [DOI] [PubMed] [Google Scholar]
  21. Palva E. T., Liljeström P., Harayama S. Cosmid cloning and transposon mutagenesis in Salmonella typhimurium using phage lambda vehicles. Mol Gen Genet. 1981;181(2):153–157. doi: 10.1007/BF00268420. [DOI] [PubMed] [Google Scholar]
  22. Pioli D., Venables W. A., Franklin F. C. D-Alanine dehydrogenase. Its role in the utilisation of alanine isomers as growth substrates by Pseudomonas aeruginosa PA01. Arch Microbiol. 1976 Nov 2;110(23):287–293. doi: 10.1007/BF00690240. [DOI] [PubMed] [Google Scholar]
  23. Raunio R. P., Straus L. D., Jenkins W. T. D-alanine oxidase from Escherichia coli: participation in the oxidation of L-alanine. J Bacteriol. 1973 Aug;115(2):567–573. doi: 10.1128/jb.115.2.567-573.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rosso G., Takashima K., Adams E. Coenzyme content of purified alanine racemase from Pseudomonas. Biochem Biophys Res Commun. 1969 Jan 6;34(1):134–140. doi: 10.1016/0006-291x(69)90539-7. [DOI] [PubMed] [Google Scholar]
  25. Shortle D., Koshland D., Weinstock G. M., Botstein D. Segment-directed mutagenesis: construction in vitro of point mutations limited to a small predetermined region of a circular DNA molecule. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5375–5379. doi: 10.1073/pnas.77.9.5375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  27. UMBARGER H. E., BROWN B. Threonine deamination in Escherichia coli. II. Evidence for two L-threonine deaminases. J Bacteriol. 1957 Jan;73(1):105–112. doi: 10.1128/jb.73.1.105-112.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Vogelstein B., Gillespie D. Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci U S A. 1979 Feb;76(2):615–619. doi: 10.1073/pnas.76.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. White R. L., Fox M. S. On the molecular basis of high negative interference. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1544–1548. doi: 10.1073/pnas.71.4.1544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wijsman H. J. The characterization of an alanine racemase mutant of Escherichia coli. Genet Res. 1972 Dec;20(3):269–277. doi: 10.1017/s001667230001380x. [DOI] [PubMed] [Google Scholar]
  31. Wild J., Klopotowski T. D-Amino acid dehydrogenase of Escherichia coli K12: positive selection of mutants defective in enzyme activity and localization of the structural gene. Mol Gen Genet. 1981;181(3):373–378. doi: 10.1007/BF00425614. [DOI] [PubMed] [Google Scholar]
  32. Wild J., Kłopotowski T. Insensitivity of D-amino acid dehydrogenase synthesis to catabolic repression in dadR mutants of Salmonella typhimurium. Mol Gen Genet. 1975;136(1):63–73. doi: 10.1007/BF00275449. [DOI] [PubMed] [Google Scholar]
  33. Wild J., Walczak W., Krajewska-Grynkiewicz K., Klopotowski T. D-amino acid dehydrogenase: the enzyme of the first step of D-histidine and D-methionine racemization in Salmonella typhimurium. Mol Gen Genet. 1974;128(2):131–146. doi: 10.1007/BF02654486. [DOI] [PubMed] [Google Scholar]
  34. Winston F., Botstein D., Miller J. H. Characterization of amber and ochre suppressors in Salmonella typhimurium. J Bacteriol. 1979 Jan;137(1):433–439. doi: 10.1128/jb.137.1.433-439.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wu T. T. A model for three-point analysis of random general transduction. Genetics. 1966 Aug;54(2):405–410. doi: 10.1093/genetics/54.2.405. [DOI] [PMC free article] [PubMed] [Google Scholar]

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