Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1988 Feb;170(2):872–876. doi: 10.1128/jb.170.2.872-876.1988

beta-Alanine auxotrophy associated with dfp, a locus affecting DNA synthesis in Escherichia coli.

E D Spitzer 1, H E Jimenez-Billini 1, B Weiss 1
PMCID: PMC210735  PMID: 3123465

Abstract

Strains containing the conditional-lethal dfp-707 mutation, which have a defect in DNA synthesis at 42 degrees C, were found to require either pantothenate or its precursor, beta-alanine, for growth at 30 degrees C. The auxotrophy and conditional lethality were corevertible. Through localized mutagenesis of the dfp-pyrE region of Escherichia coli, another mutation, dfp-1, was obtained. It conferred the auxotrophy but not the conditional lethality of dfp-707. Complementation analysis, performed with a set of plasmid-borne deletion and insertion mutations, revealed a correspondence between the complementation of each mutant phenotype and the production of the dfp gene product, previously identified as a 45-kilodalton flavoprotein. The dfp mutants had a normal level of aspartate-1-decarboxylase, which is the only enzyme known to produce beta-alanine in E. coli and which is specified by the distant panD gene. A prototrophic pseudorevertant of a dfp-1 strain was found to have retained the dfp mutation, to be genetically unstable, and to have an elevated level of aspartate-1-decarboxylase, suggesting that it had acquired a duplication of panD. It is not known what steps in pantothenate or DNA metabolism are affected by the mutant dfp product or how its flavin moiety may be involved.

Full text

PDF
874

Selected References

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

  1. Anderson R. P., Roth J. R. Gene duplication in bacteria: alteration of gene dosage by sister-chromosome exchanges. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1083–1087. doi: 10.1101/sqb.1979.043.01.120. [DOI] [PubMed] [Google Scholar]
  2. Anderson R. P., Roth J. R. Tandem genetic duplications in phage and bacteria. Annu Rev Microbiol. 1977;31:473–505. doi: 10.1146/annurev.mi.31.100177.002353. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cronan J. E., Jr Beta-alanine synthesis in Escherichia coli. J Bacteriol. 1980 Mar;141(3):1291–1297. doi: 10.1128/jb.141.3.1291-1297.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cronan J. E., Jr, Littel K. J., Jackowski S. Genetic and biochemical analyses of pantothenate biosynthesis in Escherichia coli and Salmonella typhimurium. J Bacteriol. 1982 Mar;149(3):916–922. doi: 10.1128/jb.149.3.916-922.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dunn S. D., Snell E. E. Isolation of temperature-sensitive pantothenate kinase mutants of Salmonella typhimurium and mapping of the coaA gene. J Bacteriol. 1979 Dec;140(3):805–808. doi: 10.1128/jb.140.3.805-808.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hochhauser S. J., Weiss B. Escherichia coli mutants deficient in deoxyuridine triphosphatase. J Bacteriol. 1978 Apr;134(1):157–166. doi: 10.1128/jb.134.1.157-166.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hong J. S., Ames B. N. Localized mutagenesis of any specific small region of the bacterial chromosome. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3158–3162. doi: 10.1073/pnas.68.12.3158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jackowski S., Rock C. O. Regulation of coenzyme A biosynthesis. J Bacteriol. 1981 Dec;148(3):926–932. doi: 10.1128/jb.148.3.926-932.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Lundberg L. G., Karlström O. H., Nyman P. O., Neuhard J. Isolation and characterization of the dut gene of Escherichia coli. I. Cloning in thermoinducible plasmids. Gene. 1983 Apr;22(1):115–126. doi: 10.1016/0378-1119(83)90070-7. [DOI] [PubMed] [Google Scholar]
  13. Lundberg L. G., Thoresson H. O., Karlström O. H., Nyman P. O. Nucleotide sequence of the structural gene for dUTPase of Escherichia coli K-12. EMBO J. 1983;2(6):967–971. doi: 10.1002/j.1460-2075.1983.tb01529.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. MAAS W. K., DAVIS B. D. Pantothenate studies. I. Interference by D-serine and L-aspartic acid with pantothenate synthesis in Escherichia coli. J Bacteriol. 1950 Dec;60(6):733–745. doi: 10.1128/jb.60.6.733-745.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Primerano D. A., Burns R. O. Role of acetohydroxy acid isomeroreductase in biosynthesis of pantothenic acid in Salmonella typhimurium. J Bacteriol. 1983 Jan;153(1):259–269. doi: 10.1128/jb.153.1.259-269.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rao R. N., Rogers S. G. A thermoinducible lambda phage-ColE1 plasmid chimera for the overproduction of gene products from cloned DNA segments. Gene. 1978 May;3(3):247–263. doi: 10.1016/0378-1119(78)90035-5. [DOI] [PubMed] [Google Scholar]
  17. Rao R. N., Rogers S. G. Plasmid pKC7: a vector containing ten restriction endonuclease sites suitable for cloning DNA segments. Gene. 1979 Sep;7(1):79–82. doi: 10.1016/0378-1119(79)90044-1. [DOI] [PubMed] [Google Scholar]
  18. Sancar A., Wharton R. P., Seltzer S., Kacinski B. M., Clarke N. D., Rupp W. D. Identification of the uvrA gene product. J Mol Biol. 1981 May 5;148(1):45–62. doi: 10.1016/0022-2836(81)90234-5. [DOI] [PubMed] [Google Scholar]
  19. Sevastopoulos C. G., Wehr C. T., Glaser D. A. Large-scale automated isolation of Escherichia coli mutants with thermosensitive DNA replication. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3485–3489. doi: 10.1073/pnas.74.8.3485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Spitzer E. D., Weiss B. dfp Gene of Escherichia coli K-12, a locus affecting DNA synthesis, codes for a flavoprotein. J Bacteriol. 1985 Dec;164(3):994–1003. doi: 10.1128/jb.164.3.994-1003.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Taylor A. F., Siliciano P. G., Weiss B. Cloning of the dut (deoxyuridine triphosphatase) gene of Escherichia coli. Gene. 1980 May;9(3-4):321–336. doi: 10.1016/0378-1119(90)90330-t. [DOI] [PubMed] [Google Scholar]
  22. White B. J., Hochhauser S. J., Cintron N. M., Weiss B. Genetic mapping of xthA, the structural gene for exonuclease III in Escherichia coli K-12. J Bacteriol. 1976 Jun;126(3):1082–1088. doi: 10.1128/jb.126.3.1082-1088.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Williamson J. M., Brown G. M. Purification and properties of L-Aspartate-alpha-decarboxylase, an enzyme that catalyzes the formation of beta-alanine in Escherichia coli. J Biol Chem. 1979 Aug 25;254(16):8074–8082. [PubMed] [Google Scholar]

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

RESOURCES