Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 Feb;173(3):968–977. doi: 10.1128/jb.173.3.968-977.1991

Molecular characterization of the Escherichia coli K-12 zwf gene encoding glucose 6-phosphate dehydrogenase.

D L Rowley 1, R E Wolf Jr 1
PMCID: PMC207213  PMID: 1704005

Abstract

In Escherichia coli K-12, expression of zwf, the gene for glucose 6-phosphate dehydrogenase, is coordinated with the cellular growth rate and induced by superoxide-generating agents. To initiate the study of the molecular mechanisms regulating its expression, the gene was cloned and its DNA sequence was determined. The 5' ends of zwf mRNA isolated from cells growing in glucose and acetate minimal media were mapped. The map was complex in that transcripts mapped to -45, -52, and -62, with respect to the beginning of the coding sequence. Three analytical methods were used to search the DNA sequence for putative promoters. Only one sequence for a promoter recognized by the sigma 70 form of RNA polymerase was found by all three search routines that could be aligned with a mapped transcript, indicating that the other transcripts arise by processing of the mRNA. A computer-assisted search did not reveal a thermodynamically stable long-range mRNA secondary structure that is capable of sequestering the translation initiation region, which suggests that growth-rate-dependent regulation of glucose 6-phosphate dehydrogenase level may not be carried out by a mechanism similar to the one for the gene (gnd) for 6-phosphogluconate dehydrogenase. The DNA segment between the -10 hexamer and the start point of transcription resembles the discriminator sequence of stable RNA genes, which has been implicated in stringent control and growth-rate-dependent regulation.

Full text

PDF
968

Images in this article

Selected References

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

  1. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 8. Microbiol Rev. 1990 Jun;54(2):130–197. doi: 10.1128/mr.54.2.130-197.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baker H. V., 2nd, Wolf R. E., Jr Essential site for growth rate-dependent regulation within the Escherichia coli gnd structural gene. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7669–7673. doi: 10.1073/pnas.81.24.7669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker H. V., 2nd, Wolf R. E., Jr Growth rate-dependent regulation of 6-phosphogluconate dehydrogenase level in Escherichia coli K-12: beta-galactosidase expression in gnd-lac operon fusion strains. J Bacteriol. 1983 Feb;153(2):771–781. doi: 10.1128/jb.153.2.771-781.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Banerjee S., Fraenkel D. G. Glucose-6-phosphate dehydrogenase from Escherichia coli and from a "high-level" mutant. J Bacteriol. 1972 Apr;110(1):155–160. doi: 10.1128/jb.110.1.155-160.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barcak G. J., Wolf R. E., Jr A method for unidirectional deletion mutagenesis with application to nucleotide sequencing and preparation of gene fusions. Gene. 1986;49(1):119–128. doi: 10.1016/0378-1119(86)90391-4. [DOI] [PubMed] [Google Scholar]
  6. Barcak G. J., Wolf R. E., Jr Growth-rate-dependent expression and cloning of gnd alleles from natural isolates of Escherichia coli. J Bacteriol. 1988 Jan;170(1):365–371. doi: 10.1128/jb.170.1.365-371.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Barnell W. O., Yi K. C., Conway T. Sequence and genetic organization of a Zymomonas mobilis gene cluster that encodes several enzymes of glucose metabolism. J Bacteriol. 1990 Dec;172(12):7227–7240. doi: 10.1128/jb.172.12.7227-7240.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  10. Bremer H., Berry L., Dennis P. P. Regulation of ribonucleic acid synthesis in Escherichia coli B-r: an analysis of a shift-up. II. Fraction of RNA polymerase engaged in the synthesis of stable RNA at different steady-state growth rates. J Mol Biol. 1973 Mar 25;75(1):161–179. doi: 10.1016/0022-2836(73)90536-6. [DOI] [PubMed] [Google Scholar]
  11. Camardella L., Caruso C., Rutigliano B., Romano M., Di Prisco G., Descalzi-Cancedda F. Human erythrocyte glucose-6-phosphate dehydrogenase. Identification of a reactive lysyl residue labelled with pyridoxal 5'-phosphate. Eur J Biochem. 1988 Feb 1;171(3):485–489. doi: 10.1111/j.1432-1033.1988.tb13815.x. [DOI] [PubMed] [Google Scholar]
  12. Carter-Muenchau P., Wolf R. E., Jr Growth-rate-dependent regulation of 6-phosphogluconate dehydrogenase level mediated by an anti-Shine-Dalgarno sequence located within the Escherichia coli gnd structural gene. Proc Natl Acad Sci U S A. 1989 Feb;86(4):1138–1142. doi: 10.1073/pnas.86.4.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Csonka L. N., Fraenkel D. G. Pathways of NADPH formation in Escherichia coli. J Biol Chem. 1977 May 25;252(10):3382–3391. [PubMed] [Google Scholar]
  15. Farrish E. E., Baker H. V., 2nd, Wolf R. E., Jr Different control circuits for growth rate-dependent regulation of 6-phosphogluconate dehydrogenase and protein components of the translational machinery in Escherichia coli. J Bacteriol. 1982 Nov;152(2):584–594. doi: 10.1128/jb.152.2.584-594.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  17. Fouts D., Ganguly R., Gutierrez A. G., Lucchesi J. C., Manning J. E. Nucleotide sequence of the Drosophila glucose-6-phosphate dehydrogenase gene and comparison with the homologous human gene. Gene. 1988 Mar 31;63(2):261–275. doi: 10.1016/0378-1119(88)90530-6. [DOI] [PubMed] [Google Scholar]
  18. Fraenkel D. G., Banerjee S. A mutation increasing the amount of a constitutive enzyme in Escherichia coli, glucose 6-phosphate dehydrogenase. J Mol Biol. 1971 Feb 28;56(1):183–194. doi: 10.1016/0022-2836(71)90093-3. [DOI] [PubMed] [Google Scholar]
  19. Fraenkel D. G., Banerjee S. Deletion mapping of zwf, the gene for a constitutive enzyme, glucose 6-phosphate dehydrogenase in Escherichia coli. Genetics. 1972 Aug;71(4):481–489. doi: 10.1093/genetics/71.4.481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Fraenkel D. G., Levisohn S. R. Glucose and gluconate metabolism in an Escherichia coli mutant lacking phosphoglucose isomerase. J Bacteriol. 1967 May;93(5):1571–1578. doi: 10.1128/jb.93.5.1571-1578.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Fraenkel D. G., Parola A. "Up-promoter" mutations of glucose 6-phosphate dehydrogenase in Escherichia coli. J Mol Biol. 1972 Oct 28;71(1):107–111. doi: 10.1016/0022-2836(72)90405-6. [DOI] [PubMed] [Google Scholar]
  22. Fraenkel D. G. Selection of Escherichia coli mutants lacking glucose-6-phosphate dehydrogenase or gluconate-6-phosphate dehydrogenase. J Bacteriol. 1968 Apr;95(4):1267–1271. doi: 10.1128/jb.95.4.1267-1271.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Gold L., Pribnow D., Schneider T., Shinedling S., Singer B. S., Stormo G. Translational initiation in prokaryotes. Annu Rev Microbiol. 1981;35:365–403. doi: 10.1146/annurev.mi.35.100181.002053. [DOI] [PubMed] [Google Scholar]
  24. Greenberg J. T., Demple B. A global response induced in Escherichia coli by redox-cycling agents overlaps with that induced by peroxide stress. J Bacteriol. 1989 Jul;171(7):3933–3939. doi: 10.1128/jb.171.7.3933-3939.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Grundström T., Normark S. Initiation of translation makes attenuation of ampC in E. coli dependent on growth rate. Mol Gen Genet. 1985;198(3):411–415. doi: 10.1007/BF00332931. [DOI] [PubMed] [Google Scholar]
  26. Ho Y. S., Howard A. J., Crapo J. D. Cloning and sequence of a cDNA encoding rat glucose-6-phosphate dehydrogenase. Nucleic Acids Res. 1988 Aug 11;16(15):7746–7746. doi: 10.1093/nar/16.15.7746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Jaurin B., Grundström T., Edlund T., Normark S. The E. coli beta-lactamase attenuator mediates growth rate-dependent regulation. Nature. 1981 Mar 19;290(5803):221–225. doi: 10.1038/290221a0. [DOI] [PubMed] [Google Scholar]
  28. Jeffery J., Hobbs L., Jörnvall H. Glucose-6-phosphate dehydrogenase from Saccharomyces cerevisiae: characterization of a reactive lysine residue labeled with acetylsalicylic acid. Biochemistry. 1985 Jan 29;24(3):666–671. doi: 10.1021/bi00324a019. [DOI] [PubMed] [Google Scholar]
  29. Jeffery J., Söderling-Barros J., Murray L. A., Hansen R. J., Szepesi B., Jörnvall H. Molecular diversity of glucose-6-phosphate dehydrogenase: rat enzyme structure identifies NH2-terminal segment, shows initiation from sites nonequivalent in different organisms, and establishes otherwise extensive sequence conservation. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7840–7843. doi: 10.1073/pnas.85.21.7840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kao S. M., Hassan H. M. Biochemical characterization of a paraquat-tolerant mutant of Escherichia coli. J Biol Chem. 1985 Sep 5;260(19):10478–10481. [PubMed] [Google Scholar]
  31. Kletzien R. F., Prostko C. R., Stumpo D. J., McClung J. K., Dreher K. L. Molecular cloning of DNA sequences complementary to rat liver glucose-6-phosphate dehydrogenase mRNA. Nutritional regulation of mRNA levels. J Biol Chem. 1985 May 10;260(9):5621–5624. [PubMed] [Google Scholar]
  32. 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]
  33. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. O'Neill M. C. Consensus methods for finding and ranking DNA binding sites. Application to Escherichia coli promoters. J Mol Biol. 1989 May 20;207(2):301–310. doi: 10.1016/0022-2836(89)90256-8. [DOI] [PubMed] [Google Scholar]
  36. Oliphant A. R., Struhl K. Defining the consensus sequences of E.coli promoter elements by random selection. Nucleic Acids Res. 1988 Aug 11;16(15):7673–7683. doi: 10.1093/nar/16.15.7673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Persico M. G., Viglietto G., Martini G., Toniolo D., Paonessa G., Moscatelli C., Dono R., Vulliamy T., Luzzatto L., D'Urso M. Isolation of human glucose-6-phosphate dehydrogenase (G6PD) cDNA clones: primary structure of the protein and unusual 5' non-coding region. Nucleic Acids Res. 1986 Mar 25;14(6):2511–2522. doi: 10.1093/nar/14.6.2511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Sharp P. M., Li W. H. The codon Adaptation Index--a measure of directional synonymous codon usage bias, and its potential applications. Nucleic Acids Res. 1987 Feb 11;15(3):1281–1295. doi: 10.1093/nar/15.3.1281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  43. Thomson J., Gerstenberger P. D., Goldberg D. E., Gociar E., Orozco de Silva A., Fraenkel D. G. ColE1 hybrid plasmids for Escherichia coli genes of glycolysis and the hexose monophosphate shunt. J Bacteriol. 1979 Jan;137(1):502–506. doi: 10.1128/jb.137.1.502-506.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Travers A. A. Conserved features of coordinately regulated E. coli promoters. Nucleic Acids Res. 1984 Mar 26;12(6):2605–2618. doi: 10.1093/nar/12.6.2605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  46. Wolf R. E., Jr Integration of specialized transducing bacteriophage lambda cI857 St68 h80 dgnd his by an unusual pathway promotes formation of deletions and generates a new translocatable element. J Bacteriol. 1980 May;142(2):588–602. doi: 10.1128/jb.142.2.588-602.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wolf R. E., Jr, Prather D. M., Shea F. M. Growth-rate-dependent alteration of 6-phosphogluconate dehydrogenase and glucose 6-phosphate dehydrogenase levels in Escherichia coli K-12. J Bacteriol. 1979 Sep;139(3):1093–1096. doi: 10.1128/jb.139.3.1093-1096.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wolf R. E., Jr, Shea F. M. Combined use of strain construction and affinity chromatography in the rapid, high-yield purification of 6-phosphogluconate dehydrogenase from Escherichia coli. J Bacteriol. 1979 Apr;138(1):171–175. doi: 10.1128/jb.138.1.171-175.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Zasloff M., Ginder G. D., Felsenfeld G. A new method for the purification and identification of covalently closed circular DNA molcules. Nucleic Acids Res. 1978 Apr;5(4):1139–1152. doi: 10.1093/nar/5.4.1139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Zuker M., Stiegler P. Optimal computer folding of large RNA sequences using thermodynamics and auxiliary information. Nucleic Acids Res. 1981 Jan 10;9(1):133–148. doi: 10.1093/nar/9.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES