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. 1994 Aug;176(16):4858–4864. doi: 10.1128/jb.176.16.4858-4864.1994

apbA, a new genetic locus involved in thiamine biosynthesis in Salmonella typhimurium.

D M Downs 1, L Petersen 1
PMCID: PMC196320  PMID: 7519593

Abstract

In Salmonella typhimurium, the synthesis of the pyrimidine moiety of thiamine can occur by utilization of the first five steps in de novo purine biosynthesis or independently of the pur genes through the alternative pyrimidine biosynthetic, or APB, pathway (D. M. Downs, J. Bacteriol. 174:1515-1521, 1992). We have isolated the first mutations defective in the APB pathway. These mutations define the apbA locus and map at 10.5 min on the S. typhimurium chromosome. We have cloned and sequenced the apbA gene and found it to encode a 32-kDa polypeptide whose sequence predicts an NAD/flavin adenine dinucleotide-binding pocket in the protein. The phenotypes of apbA mutants suggest that, under some conditions, the APB pathway is the sole source of the pyrimidine moiety of thiamine in wild-type S. typhimurium, and furthermore, the pur genetic background of the strain influences whether this pathway can function under aerobic and/or anaerobic growth conditions.

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Selected References

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  1. Benson N. R., Goldman B. S. Rapid mapping in Salmonella typhimurium with Mud-P22 prophages. J Bacteriol. 1992 Mar;174(5):1673–1681. doi: 10.1128/jb.174.5.1673-1681.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bochner B. R., Huang H. C., Schieven G. L., Ames B. N. Positive selection for loss of tetracycline resistance. J Bacteriol. 1980 Aug;143(2):926–933. doi: 10.1128/jb.143.2.926-933.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Castilho B. A., Olfson P., Casadaban M. J. Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons. J Bacteriol. 1984 May;158(2):488–495. doi: 10.1128/jb.158.2.488-495.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chan R. K., Botstein D., Watanabe T., Ogata Y. Specialized transduction of tetracycline resistance by phage P22 in Salmonella typhimurium. II. Properties of a high-frequency-transducing lysate. Virology. 1972 Dec;50(3):883–898. doi: 10.1016/0042-6822(72)90442-4. [DOI] [PubMed] [Google Scholar]
  5. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Downs D. M. Evidence for a new, oxygen-regulated biosynthetic pathway for the pyrimidine moiety of thiamine in Salmonella typhimurium. J Bacteriol. 1992 Mar;174(5):1515–1521. doi: 10.1128/jb.174.5.1515-1521.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Downs D. M., Roth J. R. Synthesis of thiamine in Salmonella typhimurium independent of the purF function. J Bacteriol. 1991 Oct;173(20):6597–6604. doi: 10.1128/jb.173.20.6597-6604.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Escalante-Semerena J. C., Roth J. R. Regulation of cobalamin biosynthetic operons in Salmonella typhimurium. J Bacteriol. 1987 May;169(5):2251–2258. doi: 10.1128/jb.169.5.2251-2258.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Estramareix B., Gaudry D., Thérisod M. Biosynthèse du thiazole de la thiamine chez Escherichia coli. Biochimie. 1977;59(10):857–859. doi: 10.1016/s0300-9084(77)80219-8. [DOI] [PubMed] [Google Scholar]
  10. Estramareix B., Therisod M. La tyrosine, facteur de la biosynthèse du thiazole de la thiamine chez Escherichia coli. Biochim Biophys Acta. 1972 Jul 19;273(2):275–282. [PubMed] [Google Scholar]
  11. GOLDSTEIN G. A., BROWN G. M. THE BIOSYNTHESIS OF THIAMINE. V. STUDIES CONCERNING PRECURSORS OF THE PYRIMIDINE MOIETY. Arch Biochem Biophys. 1963 Dec;103:449–452. doi: 10.1016/0003-9861(63)90436-3. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Hughes K. T., Roth J. R. Directed formation of deletions and duplications using Mud(Ap, lac). Genetics. 1985 Feb;109(2):263–282. doi: 10.1093/genetics/109.2.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kilstrup M., Meng L. M., Neuhard J., Nygaard P. Genetic evidence for a repressor of synthesis of cytosine deaminase and purine biosynthesis enzymes in Escherichia coli. J Bacteriol. 1989 Apr;171(4):2124–2127. doi: 10.1128/jb.171.4.2124-2127.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Martinez E., Bartolomé B., de la Cruz F. pACYC184-derived cloning vectors containing the multiple cloning site and lacZ alpha reporter gene of pUC8/9 and pUC18/19 plasmids. Gene. 1988 Aug 15;68(1):159–162. doi: 10.1016/0378-1119(88)90608-7. [DOI] [PubMed] [Google Scholar]
  18. Messenger L. J., Zalkin H. Glutamine phosphoribosylpyrophosphate amidotransferase from Escherichia coli. Purification and properties. J Biol Chem. 1979 May 10;254(9):3382–3392. [PubMed] [Google Scholar]
  19. Newell P. C., Tucker R. G. Biosynthesis of the pyrimidine moiety of thiamine. A new route of pyrimidine biosynthesis involving purine intermediates. Biochem J. 1968 Jan;106(1):279–287. doi: 10.1042/bj1060279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Newell P. C., Tucker R. G. New pyrimidine pathway involved in the biosynthesis of the pyrimidine of thiamine. Nature. 1967 Sep 23;215(5108):1384–1385. doi: 10.1038/2151384a0. [DOI] [PubMed] [Google Scholar]
  21. Newell P. C., Tucker R. G. Precursors of the pyrimidine moiety of thiamine. Biochem J. 1968 Jan;106(1):271–277. doi: 10.1042/bj1060271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rolfes R. J., Zalkin H. Escherichia coli gene purR encoding a repressor protein for purine nucleotide synthesis. Cloning, nucleotide sequence, and interaction with the purF operator. J Biol Chem. 1988 Dec 25;263(36):19653–19661. [PubMed] [Google Scholar]
  23. Rolfes R. J., Zalkin H. Purification of the Escherichia coli purine regulon repressor and identification of corepressors. J Bacteriol. 1990 Oct;172(10):5637–5642. doi: 10.1128/jb.172.10.5637-5642.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schmieger H. Phage P22-mutants with increased or decreased transduction abilities. Mol Gen Genet. 1972;119(1):75–88. doi: 10.1007/BF00270447. [DOI] [PubMed] [Google Scholar]
  26. Smith R. H., McAllan A. B. Nucleic acid metabolism in the ruminant. 2. Formation of microbial nucleic acids in the rumen in relation to the digestion of food nitrogen, and the fate of dietary nucleic acids. Br J Nutr. 1970 Jun;24(2):545–556. doi: 10.1079/bjn19700052. [DOI] [PubMed] [Google Scholar]
  27. Tso J. Y., Hermodson M. A., Zalkin H. Glutamine phosphoribosylpyrophosphate amidotransferase from cloned Escherichia coli purF. NH2-terminal amino acid sequence, identification of the glutamine site, and trace metal analysis. J Biol Chem. 1982 Apr 10;257(7):3532–3536. [PubMed] [Google Scholar]
  28. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  29. Vander Horn P. B., Backstrom A. D., Stewart V., Begley T. P. Structural genes for thiamine biosynthetic enzymes (thiCEFGH) in Escherichia coli K-12. J Bacteriol. 1993 Feb;175(4):982–992. doi: 10.1128/jb.175.4.982-992.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Way J. C., Davis M. A., Morisato D., Roberts D. E., Kleckner N. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene. 1984 Dec;32(3):369–379. doi: 10.1016/0378-1119(84)90012-x. [DOI] [PubMed] [Google Scholar]
  31. White R. H., Rudolph F. B. Biosynthesis of the pyrimidine moiety of thiamin in Escherichia coli: incorporation of stable isotope-labeled glycines. Biochemistry. 1979 Jun 12;18(12):2632–2636. doi: 10.1021/bi00579a031. [DOI] [PubMed] [Google Scholar]
  32. White R. H., Rudolph F. B. The origin of the nitrogen atom in the thiazole ring of thiamine in Escherichia coli. Biochim Biophys Acta. 1978 Aug 17;542(2):340–347. doi: 10.1016/0304-4165(78)90029-6. [DOI] [PubMed] [Google Scholar]
  33. White R. H. Stable isotope studies on the biosynthesis of the thiazole moiety of thiamin in Escherichia coli. Biochemistry. 1978 Sep 5;17(18):3833–3840. doi: 10.1021/bi00611a024. [DOI] [PubMed] [Google Scholar]
  34. Youderian P., Sugiono P., Brewer K. L., Higgins N. P., Elliott T. Packaging specific segments of the Salmonella chromosome with locked-in Mud-P22 prophages. Genetics. 1988 Apr;118(4):581–592. doi: 10.1093/genetics/118.4.581. [DOI] [PMC free article] [PubMed] [Google Scholar]

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