Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1989 Jun;171(6):3511–3517. doi: 10.1128/jb.171.6.3511-3517.1989

Mapping of the Escherichia coli acid glucose-1-phosphatase gene agp and analysis of its expression in vivo by use of an agp-phoA protein fusion.

E Pradel 1, P L Boquet 1
PMCID: PMC210078  PMID: 2542226

Abstract

The agp gene of Escherichia coli encodes an acid glucose-1-phosphatase, one of the numerous phosphatases optimally active between pH 4 and 6 found in the periplasmic space of this bacterium. An agp-phoA protein fusion linked to a gene conferring kanamycin resistance was inserted into the chromosome in place of agp by homologous recombination and was mapped to minute 22.6. Because the activity of glucose-1-phosphatase cannot be measured accurately in whole cells, the alkaline phosphatase activity of the agp-phoA hybrid protein was used to monitor the expression of the chromosomal agp gene. The expression of agp was subject to catabolite repression but was unaffected by the concentration of inorganic phosphate in the growth medium. The product of the agp gene was required for growth on glucose-1-phosphate as the sole carbon source, a function for which alkaline phosphatase or other acid phosphatases cannot substitute.

Full text

PDF
3511

Images in this article

3515Image
on p.3515

Selected References

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

  1. Ambudkar S. V., Larson T. J., Maloney P. C. Reconstitution of sugar phosphate transport systems of Escherichia coli. J Biol Chem. 1986 Jul 15;261(20):9083–9086. [PubMed] [Google Scholar]
  2. Beacham I. R. Periplasmic enzymes in gram-negative bacteria. Int J Biochem. 1979;10(11):877–883. doi: 10.1016/0020-711x(79)90117-4. [DOI] [PubMed] [Google Scholar]
  3. Beacham I. R., Yagil E. Genetic location of the gene (ush) specifying periplasmic uridine 5'-diphosphate glucose hydrolase (5'-nucleotidase) in Escherichia coli K-12. J Bacteriol. 1976 Oct;128(1):487–489. doi: 10.1128/jb.128.1.487-489.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boquet P. L., Manoil C., Beckwith J. Use of TnphoA to detect genes for exported proteins in Escherichia coli: identification of the plasmid-encoded gene for a periplasmic acid phosphatase. J Bacteriol. 1987 Apr;169(4):1663–1669. doi: 10.1128/jb.169.4.1663-1669.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brickman E., Beckwith J. Analysis of the regulation of Escherichia coli alkaline phosphatase synthesis using deletions and phi80 transducing phages. J Mol Biol. 1975 Aug 5;96(2):307–316. doi: 10.1016/0022-2836(75)90350-2. [DOI] [PubMed] [Google Scholar]
  6. Burns D. M., Beacham I. R. Nucleotide sequence and transcriptional analysis of the E. coli ushA gene, encoding periplasmic UDP-sugar hydrolase (5'-nucleotidase): regulation of the ushA gene, and the signal sequence of its encoded protein product. Nucleic Acids Res. 1986 May 27;14(10):4325–4342. doi: 10.1093/nar/14.10.4325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. DAVIS B. D., MINGIOLI E. S. Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950 Jul;60(1):17–28. doi: 10.1128/jb.60.1.17-28.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dassa E., Boquet P. L. Identification of the gene appA for the acid phosphatase (pH optimum 2.5) of Escherichia coli. Mol Gen Genet. 1985;200(1):68–73. doi: 10.1007/BF00383314. [DOI] [PubMed] [Google Scholar]
  9. Dassa E., Cahu M., Desjoyaux-Cherel B., Boquet P. L. The acid phosphatase with optimum pH of 2.5 of Escherichia coli. Physiological and Biochemical study. J Biol Chem. 1982 Jun 25;257(12):6669–6676. [PubMed] [Google Scholar]
  10. Dietz G. W., Heppel L. A. Studies on the uptake of hexose phosphates. 3. Mechanism of uptake of glucose 1-phosphate in Escherichia coli. J Biol Chem. 1971 May 10;246(9):2891–2897. [PubMed] [Google Scholar]
  11. Dietz G. W., Heppel L. A. Studies on the uptake of hexose phosphates. I. 2-Deoxyglucose and 2-deoxyglucose 6-phosphate. J Biol Chem. 1971 May 10;246(9):2881–2884. [PubMed] [Google Scholar]
  12. Dietz G. W., Heppel L. A. Studies on the uptake of hexose phosphates. II. The induction of the glucose 6-phosphate transport system by exogenous but not by endogenously formed glucose 6-phosphate. J Biol Chem. 1971 May 10;246(9):2885–2890. [PubMed] [Google Scholar]
  13. Dietz G. W., Jr The hexose phosphate transport system of Escherichia coli. Adv Enzymol Relat Areas Mol Biol. 1976;44:237–259. doi: 10.1002/9780470122891.ch7. [DOI] [PubMed] [Google Scholar]
  14. Dvorak H. F., Brockman R. W., Heppel L. A. Purification and properties of two acid phosphatase fractions isolated from osmotic shock fluid of Escherichia coli. Biochemistry. 1967 Jun;6(6):1743–1751. doi: 10.1021/bi00858a024. [DOI] [PubMed] [Google Scholar]
  15. Kadner R. J. Genetic Control of the Transport of Hexose Phosphates in Escherichia coli: Mapping of the uhp Locus. J Bacteriol. 1973 Nov;116(2):764–770. doi: 10.1128/jb.116.2.764-770.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kadner R. J., Shattuck-Eidens D. M. Genetic control of the hexose phosphate transport system of Escherichia coli: mapping of deletion and insertion mutations in the uhp region. J Bacteriol. 1983 Sep;155(3):1052–1061. doi: 10.1128/jb.155.3.1052-1061.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kadner R. J., Winkler H. H. Isolation and characterization of mutations affecting the transport of hexose phosphates in Escherichia coli. J Bacteriol. 1973 Feb;113(2):895–900. doi: 10.1128/jb.113.2.895-900.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
  19. Manoil C., Beckwith J. A genetic approach to analyzing membrane protein topology. Science. 1986 Sep 26;233(4771):1403–1408. doi: 10.1126/science.3529391. [DOI] [PubMed] [Google Scholar]
  20. Manoil C., Beckwith J. TnphoA: a transposon probe for protein export signals. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129–8133. doi: 10.1073/pnas.82.23.8129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Michaelis S., Inouye H., Oliver D., Beckwith J. Mutations that alter the signal sequence of alkaline phosphatase in Escherichia coli. J Bacteriol. 1983 Apr;154(1):366–374. doi: 10.1128/jb.154.1.366-374.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pogell B. M., Maity B. R., Frumkin S., Shapiro S. Induction of an active transport system for glucose 6-phosphate in Escherichia coli. Arch Biochem Biophys. 1966 Sep 26;116(1):406–415. doi: 10.1016/0003-9861(66)90047-6. [DOI] [PubMed] [Google Scholar]
  23. Pradel E., Boquet P. L. Acid phosphatases of Escherichia coli: molecular cloning and analysis of agp, the structural gene for a periplasmic acid glucose phosphatase. J Bacteriol. 1988 Oct;170(10):4916–4923. doi: 10.1128/jb.170.10.4916-4923.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. ROGERS D., REITHEL F. J. Acid phosphatases of Escherichia coli. Arch Biochem Biophys. 1960 Jul;89:97–104. doi: 10.1016/0003-9861(60)90018-7. [DOI] [PubMed] [Google Scholar]
  25. Rephaeli A. W., Artenstein I. R., Saier M. H., Jr Physiological function of periplasmic hexose phosphatase in Salmonella typhimurium. J Bacteriol. 1980 Mar;141(3):1474–1477. doi: 10.1128/jb.141.3.1474-1477.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sarthy A., Michaelis S., Beckwith J. Deletion map of the Escherichia coli structural gene for alkaline phosphatase, phoA. J Bacteriol. 1981 Jan;145(1):288–292. doi: 10.1128/jb.145.1.288-292.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Shattuck-Eidens D. M., Kadner R. J. Molecular cloning of the uhp region and evidence for a positive activator for expression of the hexose phosphate transport system of Escherichia coli. J Bacteriol. 1983 Sep;155(3):1062–1070. doi: 10.1128/jb.155.3.1062-1070.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. TORRIANI A. Influence of inorganic phosphate in the formation of phosphatases by Escherichia coli. Biochim Biophys Acta. 1960 Mar 11;38:460–469. doi: 10.1016/0006-3002(60)91281-6. [DOI] [PubMed] [Google Scholar]
  30. Touati E., Dassa E., Boquet P. L. Pleiotropic mutations in appR reduce pH 2.5 acid phosphatase expression and restore succinate utilisation in CRP-deficient strains of Escherichia coli. Mol Gen Genet. 1986 Feb;202(2):257–264. doi: 10.1007/BF00331647. [DOI] [PubMed] [Google Scholar]
  31. Weston L. A., Kadner R. J. Identification of uhp polypeptides and evidence for their role in exogenous induction of the sugar phosphate transport system of Escherichia coli K-12. J Bacteriol. 1987 Aug;169(8):3546–3555. doi: 10.1128/jb.169.8.3546-3555.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weston L. A., Kadner R. J. Role of uhp genes in expression of the Escherichia coli sugar-phosphate transport system. J Bacteriol. 1988 Aug;170(8):3375–3383. doi: 10.1128/jb.170.8.3375-3383.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Winkler H. H. A hexose-phosphate transport system in Escherichia coli. Biochim Biophys Acta. 1966 Mar 28;117(1):231–240. doi: 10.1016/0304-4165(66)90170-x. [DOI] [PubMed] [Google Scholar]
  34. Winkler H. H. Compartmentation in the induction of the hexose-6-phosphate transport system of Escherichia coli. J Bacteriol. 1970 Feb;101(2):470–475. doi: 10.1128/jb.101.2.470-475.1970. [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]
  36. von HOFSTEN, PORATH J. Purification and some properties of an acid phosphatase from Escherichia coli. Biochim Biophys Acta. 1962 Oct 8;64:1–12. doi: 10.1016/0006-3002(62)90754-0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES