Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Aug;169(8):3546–3555. doi: 10.1128/jb.169.8.3546-3555.1987

Identification of uhp polypeptides and evidence for their role in exogenous induction of the sugar phosphate transport system of Escherichia coli K-12.

L A Weston, R J Kadner
PMCID: PMC212431  PMID: 3038843

Abstract

Cells of Escherichia coli possess a transport system that catalyzes the accumulation, in unaltered form, of a variety of sugar phosphates. Induction of the transport activity occurs in response to external glucose 6-phosphate and does not require detectable entry of this inducer. To define the genes that encode the Uhp transport system and those that mediate its exogenous induction, transposon insertions were isolated and mapped within a 6.5-kilobase HindIII-BamHI fragment that carries the entire uhp region. The transposon insertions were transferred by homologous recombination onto the chromosome to test their effect on Uhp expression when all genes were present in single copy number. The complementation behavior of plasmids carrying the insertions or subcloned fragments of the region was compared with their polypeptide coding capacity in maxicells. These studies defined three uhp regulatory genes (uhpABC), all of which are necessary for expression of the uhpT gene, which encodes the transporter. The products of uhpB and uhpC are not required when uhpA is present on a multicopy plasmid. The four genes, uhpA, uhpB, uhpC, and uhpT, are transcribed in the same direction, and their products have apparent molecular weights of 25,000, 48,000, 20,000, and 38,000, respectively. The UhpB and UhpT polypeptides are associated with the membrane fraction. These results led to a model of regulation in which the UhpB and UhpC regulatory proteins prevent the ability of UhpA to activate transcription of the uhpT gene under noninducing conditions.

Full text

PDF
3547

Images in this article

3552Image
on p.3552
3552Image
on p.3552
3553Image
on p.3553
3554Image
on p.3554

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. 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]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clarke L., Carbon J. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell. 1976 Sep;9(1):91–99. doi: 10.1016/0092-8674(76)90055-6. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Eidels L., Rick P. D., Stimler N. P., Osborn M. J. Transport of D-arabinose-5-phosphate and D-sedoheptulose-7-phosphate by the hexose phosphate transport system of Salmonella typhimurium. J Bacteriol. 1974 Jul;119(1):138–143. doi: 10.1128/jb.119.1.138-143.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Essenberg R. C., Kornberg H. L. Location of the gene specifying hexose phosphate transport (uhp) on the chromosome of Escherichia coli. J Gen Microbiol. 1977 Mar;99(1):157–169. doi: 10.1099/00221287-99-1-157. [DOI] [PubMed] [Google Scholar]
  9. Fecker L., Braun V. Cloning and expression of the fhu genes involved in iron(III)-hydroxamate uptake by Escherichia coli. J Bacteriol. 1983 Dec;156(3):1301–1314. doi: 10.1128/jb.156.3.1301-1314.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Friedrich M. J., Kadner R. J. Nucleotide sequence of the uhp region of Escherichia coli. J Bacteriol. 1987 Aug;169(8):3556–3563. doi: 10.1128/jb.169.8.3556-3563.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Greener A., Hill C. W. Identification of a novel genetic element in Escherichia coli K-12. J Bacteriol. 1980 Oct;144(1):312–321. doi: 10.1128/jb.144.1.312-321.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guth A., Engel R., Tropp B. E. Uptake of glycerol 3-phosphate and some of its analogs by the hexose phosphate transport system of Escherichia coli. J Bacteriol. 1980 Jul;143(1):538–539. doi: 10.1128/jb.143.1.538-539.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Guyer M. S. The gamma delta sequence of F is an insertion sequence. J Mol Biol. 1978 Dec 15;126(3):347–365. doi: 10.1016/0022-2836(78)90045-1. [DOI] [PubMed] [Google Scholar]
  14. Inouye M., Guthrie J. P. A mutation which changes a membrane protein of E. coli. Proc Natl Acad Sci U S A. 1969 Nov;64(3):957–961. doi: 10.1073/pnas.64.3.957. [DOI] [PMC free article] [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. Kahan F. M., Kahan J. S., Cassidy P. J., Kropp H. The mechanism of action of fosfomycin (phosphonomycin). Ann N Y Acad Sci. 1974 May 10;235(0):364–386. doi: 10.1111/j.1749-6632.1974.tb43277.x. [DOI] [PubMed] [Google Scholar]
  19. Larson T. J., Schumacher G., Boos W. Identification of the glpT-encoded sn-glycerol-3-phosphate permease of Escherichia coli, an oligomeric integral membrane protein. J Bacteriol. 1982 Dec;152(3):1008–1021. doi: 10.1128/jb.152.3.1008-1021.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. LeBlanc G., Rimon G., Kaback H. R. Glucose 6-phosphate transport in membrane vesicles isolated from Escherichia coli: effect of imposed electrical potential and pH gradient. Biochemistry. 1980 May 27;19(11):2522–2528. doi: 10.1021/bi00552a034. [DOI] [PubMed] [Google Scholar]
  21. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [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. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Wek R. C., Hauser C. A., Hatfield G. W. The nucleotide sequence of the ilvBN operon of Escherichia coli: sequence homologies of the acetohydroxy acid synthase isozymes. Nucleic Acids Res. 1985 Jun 11;13(11):3995–4010. doi: 10.1093/nar/13.11.3995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Winans S. C., Elledge S. J., Krueger J. H., Walker G. C. Site-directed insertion and deletion mutagenesis with cloned fragments in Escherichia coli. J Bacteriol. 1985 Mar;161(3):1219–1221. doi: 10.1128/jb.161.3.1219-1221.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Winkler H. H. Energy coupling of the hexose phosphate transport system in Escherichia coli. J Bacteriol. 1973 Oct;116(1):203–209. doi: 10.1128/jb.116.1.203-209.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES