1887

Abstract

Previous work has resulted in the isolation of several mutant glucose permeases (II or PtsG) of the phosphotransferase system (PTS) with altered N-terminal amphipathic leader sequences. The mutations were reported to (1) broaden permease substrate specificity, (2) promote facilitated diffusion of some sugars and (3) increase gene transcription. Detailed biochemical analyses were conducted, showing that one such mutant (V12F-II) (1) contains dramatically increased amounts of II, (2) displays correspondingly increased phosphorylation and transport activities, (3) shows increased utilization of several metabolizable sugars and (4) shows decreased susceptibility to detergent activation. These results are interpreted as suggesting that the V12F substitution in the N-terminal amphipathic leader sequence of II alters the facility with which the permease is integrated into the membrane. Consequent changes in conformation alter its catalytic properties and increase its affinity for the pleiotropic transcriptional repressor, Mlc. These changes together are proposed to promote transcription of the gene and account for the observed phenotypic changes.

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2003-03-01
2024-04-19
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References

  1. Aboulwafa M., Saier M. H. Jr 2002; Dependency of sugar transport and phosphorylation by the phospho enol pyruvate-dependent phosphotransferase system on membranous phosphatidyl glycerol in Escherichia coli : studies with a pgsA mutant lacking phosphatidyl glycerophosphate synthase. Res Microbiol 153:667–677
    [Google Scholar]
  2. Ashworth J. M., Kornberg H. L. 1966; The anaplerotic fixation of carbon dioxide by Escherichia coli . Proc R Soc Lond Ser B 65:179–188
    [Google Scholar]
  3. Bogdanov M., Dowhan W. 1999; Lipid-assisted protein folding. J Biol Chem 274:36827–36830
    [Google Scholar]
  4. Bogdanov M., Heacock P. N., Dowhan W. 2002; A polytopic membrane protein displays a reversible topology dependent on membrane lipid composition. EMBO J 21:2107–2116
    [Google Scholar]
  5. Gegley G. S., Warner K. A., Arents J. C., Postma P. W., Jacobson G. R. 1996; Isolation and characterization of a mutation that alters the substrate specificity of the Escherichia coli glucose permease. J Bacteriol 178:940–942
    [Google Scholar]
  6. Kornberg H. L., Lambourne L. T. M., Sproul A. A. 2000; Facilitated diffusion of fructose via the phospho enol pyruvate/glucose phosphotransferase system of Escherichia coli . Proc Natl Acad Sci U S A 97:1808–1812
    [Google Scholar]
  7. Lee S.-J., Boos W., Bouché J.-P., Plumbridge J. 2000; Signal transduction between a membrane-bound transporter, PtsG, and a soluble transcription factor, Mlc, of Escherichia coli . EMBO J 19:5353–5361
    [Google Scholar]
  8. Nam T. W., Cho S. H., Shin D. 8 other authors 2001; The Escherichia coli glucose transporter enzyme IICB(Glc) recruits the global repressor Mlc. EMBO J 20:491–498
    [Google Scholar]
  9. Notley-McRobb L., Ferenci T. 2000; Substrate specificity and signal transduction pathways in the glucose-specific Enzyme II (EIIGlc) component of the Escherichia coli phosphotransferase system. J Bacteriol 182:4437–4442
    [Google Scholar]
  10. Plumbridge J. 1999; Expression of the phosphotransferase system both mediates and is mediated by Mlc regulation in Escherichia coli . Mol Microbiol 33:260–273
    [Google Scholar]
  11. Plumbridge J. 2000; A mutation which affects both the specificity of PtsG sugar transport and the regulation of ptsG expression by Mlc in Escherichia coli . Microbiology 146:2655–2663
    [Google Scholar]
  12. Plumbridge J. 2001; Regulation of PTS gene expression by the homologous transcriptional regulators, Mlc and NagC, in Escherichia coli (or how two similar repressors can behave differently. J Mol Microbiol Biotechnol 3:371–380
    [Google Scholar]
  13. Reidl J., Boos W. 1991; The malX malY operon of Escherichia coli encodes a novel Enzyme II of the phosphotransferase system recognizing glucose and maltose and an enzyme abolishing the endogenous induction of the maltose system. J Bacteriol 173:4862–4876
    [Google Scholar]
  14. Saier M. H. Jr 1989; Protein phosphorylation and allosteric control of inducer exclusion and catabolite repression by the bacterial phospho enol pyruvate : sugar phosphotransferase system. Microbiol Rev 53:109–120
    [Google Scholar]
  15. Saier M. H. Jr, McCaldon P. 1988; Statistical and functional analyses of viral and cellular proteins with N-terminal amphipathic α -helices with large hydrophobic moments. Importance to macromolecular recognition and organellar targeting. J Bacteriol 170:2296–2300
    [Google Scholar]
  16. Saier M. H. Jr, Reizer J. 1994; The bacterial phosphotransferase system: new frontiers 30 years later. Mol Microbiol 13:755–764
    [Google Scholar]
  17. Saier M. H. Jr, Young W. S. III, Roseman S. 1971; Utilization and transport of hexoses by mutant strains of Salmonella typhimurium lacking enzyme I of the phospho enol pyruvate-dependent phosphotransferase system. J Biol Chem 246:5838–5840
    [Google Scholar]
  18. Saier M. H. Jr, Bromberg F. G., Roseman S. 1973; Characterization of constitutive galactose permease mutants in Salmonella typhimurium . J Bacteriol 113:512–514
    [Google Scholar]
  19. Saier M. H. Jr, Cox D. F., Moczydlowski E. G. 1977a; Sugar phosphate : sugar transphosphorylation coupled to exchange group translocation catalyzed by the Enzyme II complexes of the phospho enol pyruvate : sugar phosphotransferase system in membrane vesicles of Escherichia coli . J Biol Chem 252:8908–8916
    [Google Scholar]
  20. Saier M. H. Jr, Feucht B. U., Mora W. K. 1977b; Sugar phosphate : sugar transphosphorylation and exchange group translocation catalyzed by the Enzyme II complexes of the bacterial phospho enol pyruvate : sugar phosphotransferase system. J Biol Chem 252:8899–8907
    [Google Scholar]
  21. Saier M. H. Jr, Newman M. J., Rephaeli A. W. 1977c; Properties of a phospho enol pyruvate : mannitol phosphotransferase system in Spirochaeta aurantia . J Biol Chem 252:8890–8898
    [Google Scholar]
  22. Saier M. H. Jr, Yamada M., Suda K., Erni B., Rak B., Lengeler J., Stewart G. C., Waygood E. B., Rapoport G. 1988; Bacterial proteins with N-terminal leader sequences resembling mitochondrial targeting sequences of eukaryotes. Biochemie 70:1743–1748
    [Google Scholar]
  23. Saier M. H. Jr, Schnierow B., Yamada Y., Daniels G. A. 1989a; Structures, evolution and membrane insertion of the integral membrane permease proteins of the bacterial phosphotransferase system. In Highlights of Modern Biochemistry vol. 1 pp  771–780 Edited by Kotyk A., Skoda J., Paces V., Kostk V. Zeist: VSP International Science;
    [Google Scholar]
  24. Saier M. H. Jr, Werner P., Müller M. 1989b; Insertion of proteins into bacterial membranes: mechanism, characteristics and comparisons with the eukaryotic process. Microbiol Rev 53:333–366
    [Google Scholar]
  25. Sproul A. A., Lambourne L. T., Jean-Jacques D. J., Kornberg H. L. 2001; Genetic control of manno(fructo)kinase activity in Escherichia coli . Proc Natl Acad Sci U S A 98:15257–15259
    [Google Scholar]
  26. Tamm L. K., Tomich J. M., Saier M. H. Jr 1989; Membrane incorporation and induction of secondary structure of synthetic peptides corresponding to the N-terminal signal sequences of the glucitol and mannitol permeases of Escherichia coli . J Biol Chem 264:2587–2592
    [Google Scholar]
  27. Tanaka Y., Kimata K., Aiba H. 2000; A novel regulatory role of glucose transporter of Escherichia coli : membrane sequestration of a global repressor Mlc. EMBO J 19:5344–5352
    [Google Scholar]
  28. Yamada Y., Chang Y.-Y., Daniels G. A., Wu L.-F., Tomich J. M., Yamada M., Saier M. H. Jr 1991; Insertion of the mannitol permease into the membrane of Escherichia coli . Possible involvement of an N-terminal amphiphilic sequence. J Biol Chem 266:17863–17871
    [Google Scholar]
  29. Zeppenfeld T., Larisch C., Lengeler J. W., Jahreis K. 2000; Glucose transporter mutants of Escherichia coli K-12 with changes in substrate recognition of IICBGlc and induction behaviour of the ptsG gene. J Bacteriol 182:4443–4452
    [Google Scholar]
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