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. 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.

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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]

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