Skip to main content
PMC home page
Genetics logoLink to Genetics
. 1996 May;143(1):37–44. doi: 10.1093/genetics/143.1.37

Genetic Analysis of Metabolic Crosstalk and Its Impact on Thiamine Synthesis in Salmonella Typhimurium

L Petersen 1, J Enos-Berlage 1, D M Downs 1
PMCID: PMC1207269  PMID: 8722760

Abstract

The first five steps in de novo purine biosynthesis are involved in the formation of the 4-amino-5-hydroxymethyl-2-methyl pyrimidine (HMP) moiety of thiamine. We show here that the first enzyme in de novo purine biosynthesis, PurF, is required for thiamine synthesis during aerobic growth on some but not other carbon sources. We show that PurF-independent thiamine synthesis depends on the recently described alternative pyrimidine biosynthetic (APB) pathway. Null mutations in zwf (encoding glucose-6-P dehydogenase), gnd (encoding gluconate-6-P dehydrogenase), purE (encoding aminoimidazole ribo-nucleotide carboxylase), and purR (encoding a regulator of gene expression) were found to affect the function of the APB pathway. A model is presented to account for the involvement of these gene products in thiamine biosynthesis via the APB pathway. Results presented herein demonstrate that function of the APB pathway can be prevented either by blocking intermediate formation or by diverting intermediate(s) from the pathway. Strong genetic evidence supports the conclusion that aminoimidazole ribotide (AIR) is an intermediate in the APB pathway.

Full Text

The Full Text of this article is available as a PDF (895.7 KB).

Selected References

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

  1. Balch W. E., Wolfe R. S. New approach to the cultivation of methanogenic bacteria: 2-mercaptoethanesulfonic acid (HS-CoM)-dependent growth of Methanobacterium ruminantium in a pressureized atmosphere. Appl Environ Microbiol. 1976 Dec;32(6):781–791. doi: 10.1128/aem.32.6.781-791.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. 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]
  5. Downs D. M., Petersen L. apbA, a new genetic locus involved in thiamine biosynthesis in Salmonella typhimurium. J Bacteriol. 1994 Aug;176(16):4858–4864. doi: 10.1128/jb.176.16.4858-4864.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Enos-Berlage J. L., Downs D. M. Involvement of the oxidative pentose phosphate pathway in thiamine biosynthesis in Salmonella typhimurium. J Bacteriol. 1996 Mar;178(5):1476–1479. doi: 10.1128/jb.178.5.1476-1479.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Estramareix B., David S. Conversion of 5-aminoimidazole ribotide to the pyrimidine of thiamin in enterobacteria: study of the pathway with specifically labeled samples of riboside. Biochim Biophys Acta. 1990 Aug 17;1035(2):154–160. doi: 10.1016/0304-4165(90)90110-i. [DOI] [PubMed] [Google Scholar]
  9. Gutnick D., Calvo J. M., Klopotowski T., Ames B. N. Compounds which serve as the sole source of carbon or nitrogen for Salmonella typhimurium LT-2. J Bacteriol. 1969 Oct;100(1):215–219. doi: 10.1128/jb.100.1.215-219.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kupor S. R., Fraenkel D. G. Glucose metabolism in 6 phosphogluconolactonase mutants of Escherichia coli. J Biol Chem. 1972 Mar 25;247(6):1904–1910. [PubMed] [Google Scholar]
  11. LaRossa R. A., Van Dyk T. K., Smulski D. R. Toxic accumulation of alpha-ketobutyrate caused by inhibition of the branched-chain amino acid biosynthetic enzyme acetolactate synthase in Salmonella typhimurium. J Bacteriol. 1987 Apr;169(4):1372–1378. doi: 10.1128/jb.169.4.1372-1378.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Meng L. M., Kilstrup M., Nygaard P. Autoregulation of PurR repressor synthesis and involvement of purR in the regulation of purB, purC, purL, purMN and guaBA expression in Escherichia coli. Eur J Biochem. 1990 Jan 26;187(2):373–379. doi: 10.1111/j.1432-1033.1990.tb15314.x. [DOI] [PubMed] [Google Scholar]
  13. Nasoff M. S., Baker H. V., 2nd, Wolf R. E., Jr DNA sequence of the Escherichia coli gene, gnd, for 6-phosphogluconate dehydrogenase. Gene. 1984 Mar;27(3):253–264. doi: 10.1016/0378-1119(84)90070-2. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. 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]
  17. Rolfes R. J., Zalkin H. Autoregulation of Escherichia coli purR requires two control sites downstream of the promoter. J Bacteriol. 1990 Oct;172(10):5758–5766. doi: 10.1128/jb.172.10.5758-5766.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. SUSSMAN R., JACOB F. [On a thermosensitive repression system in the Escherichia coli lambda bacteriophage]. C R Hebd Seances Acad Sci. 1962 Feb 19;254:1517–1519. [PubMed] [Google Scholar]
  21. 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]
  22. Shattuck-Eidens D. M., Kadner R. J. Exogenous induction of the Escherichia coli hexose phosphate transport system defined by uhp-lac operon fusions. J Bacteriol. 1981 Oct;148(1):203–209. doi: 10.1128/jb.148.1.203-209.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Steiert J. G., Rolfes R. J., Zalkin H., Stauffer G. V. Regulation of the Escherichia coli glyA gene by the purR gene product. J Bacteriol. 1990 Jul;172(7):3799–3803. doi: 10.1128/jb.172.7.3799-3803.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Turnbough C. L., Jr, Bochner B. R. Toxicity of the pyrimidine biosynthetic pathway intermediate carbamyl aspartate in Salmonella typhimurium. J Bacteriol. 1985 Aug;163(2):500–505. doi: 10.1128/jb.163.2.500-505.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Wilson R. L., Stauffer L. T., Stauffer G. V. Roles of the GcvA and PurR proteins in negative regulation of the Escherichia coli glycine cleavage enzyme system. J Bacteriol. 1993 Aug;175(16):5129–5134. doi: 10.1128/jb.175.16.5129-5134.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES