Archives of Microbiology

, Volume 189, Issue 6, pp 597–604 | Cite as

Escherichia coli tat mutant strains are able to transport maltose in the absence of an active malE gene

Original Paper
First Online:
Received:
Revised:
Accepted:

Abstract

The twin-arginine transport (Tat) system is a prokaryotic protein transport system. Escherichia coli mutants in this pathway show a defect in cell separation during cell division, resulting in destabilization and permeability of the outer membrane. Maltose uptake is catalysed by a membrane-bound transporter of the ATP binding cassette (ABC) superfamily, where MalE is the essential periplasmic binding protein component. Here, we report that tat mutants are unexpectedly able to transport maltose in the absence of malE. This observation is specific to the MalE component since co-inactivation of malF, which encodes one of the channel components of the transporter, completely abolishes maltose transport even when the Tat system is inactivated. Genetic repair of the outer membrane leaky phenotype of the tat mutant strain re-established the absolute requirement for MalE in maltose uptake. In addition, we demonstrate that phenotypic repair of the outer membrane defect of the tat strain can also be achieved chemically by the inclusion of high concentrations of calcium or magnesium in the growth medium.

Keywords

Protein transport Tat pathway Twin-arginine signal peptide ABC transporter Periplasmic binding protein Outer membrane 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

We thank Ms Jenny Graham for her assistance with preliminary experiments, Dr Matthew Hicks for his help in constructing the apramycin-marked tatB mutant, and Drs Ben Berks and Bérengère Ize for helpful discussion. This work is funded by the BBSRC grant BB/C516195/1. TP is a MRC Senior Non-Clinical Research Fellow and FS is a Royal Society University Research Fellow.

References

  1. Berks BC, Palmer T, Sargent F (2003) The Tat protein translocation pathway and its role in microbial physiology. Adv Microb Physiol 47:187–254PubMedCrossRefGoogle Scholar
  2. Bernhardt TG, de Boer PA (2003) The Escherichia coli amidase AmiC is a periplasmic septal ring component exported via the twin-arginine transport pathway. Mol Microbiol 48:1171–1182PubMedCrossRefGoogle Scholar
  3. Blaudeck N, Kreutzenbeck P, Freudl R, Sprenger GA (2003) Genetic analysis of pathway specificity during posttranslational protein translocation across the Escherichia coli plasma membrane. J Bacteriol 185:2811–2819PubMedCrossRefGoogle Scholar
  4. Boos W, Shuman H (1998) Maltose/maltodextrin system of Escherichia coli: transport, metabolism and regulation. Microbiol Mol Biol Rev 62:204–229PubMedGoogle Scholar
  5. Buchanan G, de Leeuw E, Stanley NR, Wexler M, Berks BC, Sargent F, Palmer T (2002). doi: 10.1046/j.1365-2958.2002.02853.x
  6. Casadaban MJ, Cohen SN (1979) Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci USA 76:4530–4533PubMedCrossRefGoogle Scholar
  7. Chatterjee AK, Ross H, Sanderson KE (1976) Leakage of periplasmic from lipopolysaccharide defective mutants of Salmonella typhimurium. Can J Microbiol 22:1549–1560PubMedGoogle Scholar
  8. Cline K, Theg SM (2007) The Sec and Tat protein translocation pathways. In: Dalbey RE, Kohler C (eds) The Enzymes. ISBN: 978-0-12-373916-2Google Scholar
  9. Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 97:6640–6645PubMedCrossRefGoogle Scholar
  10. Fekkes P, Driessen AJM (1999) Protein targeting to the bacterial cytoplasmic membrane. Microbiol Mol Biol Rev 63:161–173PubMedGoogle Scholar
  11. De Pina K, Desjardin V, Mandrand-Berthelot MA, Giordano G, Wu LF (1999) Isolation and characterization of the nikR gene encoding a nickel-responsive regulator in Escherichia coli. J Bacteriol 181:670–674PubMedGoogle Scholar
  12. Franklin NC (1969) Mutation in the galU gene of E. coli blocks phage P1 infection. Virology 38:189–191PubMedCrossRefGoogle Scholar
  13. Galanos C, Lüderitz O (1975) Electrodialysis of lipopolysaccharides and their conversion to uniform salt forms. Eur J Biochem 54:603–610PubMedCrossRefGoogle Scholar
  14. Gust B, Challis GL, Fowler K, Kieser T, Chater KF (2003) PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin. Proc Natl Acad Sci USA 100:1541–1546PubMedCrossRefGoogle Scholar
  15. Hamilton CM, Aldea M, Washburn BK, Babitzke P, Kushner SR (1989) New method for generating deletions and gene replacements in Escherichia coli. J Bacteriol 171:4617–4622PubMedGoogle Scholar
  16. Ize B, Stanley NR, Buchanan G, Palmer T (2003) Role of the Escherichia coli Tat pathway in outer membrane integrity. Mol Microbiol 48:1183–1193PubMedCrossRefGoogle Scholar
  17. Ize B, Porcelli I, Lucchini S, Hinton JC, Berks BC, Palmer T (2004) Novel phenotypes of Escherichia coli tat mutants revealed by global gene expression and phenotypic analysis. J Biol Chem 279:47543–47554PubMedCrossRefGoogle Scholar
  18. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  19. Miller JH (1992) A short course in bacterial genetics: a laboratory manual and handbook for Escherichia coli and related bacteria. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  20. Meloni S, Rey L, Sidler S, Imperial J, Ruiz-Argueso T, Palacios JM (2003) The twin-arginine translocation (Tat) system is essential for Rhizobium-legume symbiosis. Mol Microbiol 48:1195–1207PubMedCrossRefGoogle Scholar
  21. Navarro C, Wu LF, Mandrand-Berthelot MA (1993) The nik operon of Escherichia coli encodes a periplasmic binding-protein-dependent transport system for nickel. Mol Microbiol 9:1181–1191PubMedCrossRefGoogle Scholar
  22. Nikaido H, Vaara M (1985) Molecular basis of bacterial outer membrane permeability. Microbiol Rev 49:1–32PubMedGoogle Scholar
  23. Nikaido H, Bavoil P, Hirota Y (1977) Outer membranes of gram-negative bacteria. XV. Transmembrane diffusion rates in lipoprotein-deficient mutants of Escherichia coli. J Bacteriol 132:1045–1047PubMedGoogle Scholar
  24. Ochsner UA, Snyder A, Vasil AI, Vasil ML (2002) Effects of the twin-arginine translocase on secretion of virulence factors, stress response, and pathogenesis. Proc Natl Acad Sci USA 99:8312–8317PubMedCrossRefGoogle Scholar
  25. Oldham MK, Khare D, Quiochio FA, Davidson AL, Chen J (2007) Crystal structure of a catalytic intermediate of the maltose transporter. Nature 450:515–522PubMedCrossRefGoogle Scholar
  26. Pederson PL (2005) Transport ATPases: structure, motors, mechanism and medicine: a brief overview. J Bioenerg Biomembr 37:349–357CrossRefGoogle Scholar
  27. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory Manual. 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  28. Sargent F, Bogsch EG, Stanley NR, Wexler M, Robinson C, Berks BC, Palmer T (1998) Overlapping functions of components of a bacterial Sec-independent protein export pathway. EMBO J 17:3640–3650PubMedCrossRefGoogle Scholar
  29. Sargent F, Stanley NR, Berks BC, Palmer T (1999) Sec-independent protein translocation in Escherichia coli: a distinct and pivotal role for the TatB protein. J Biol Chem 274:36073–36082PubMedCrossRefGoogle Scholar
  30. Sawers G (1994) The hydrogenases and formate dehydrogenases of Escherichia coli. Antonie Van Leeuwenhoek 66:57–88PubMedCrossRefGoogle Scholar
  31. Shuman HA (1982) Periplasmic binding protein independent active transport of maltose in a mutant of Escherichia coli K-12: evidence for a substrate recognition site in the cytoplasmic membrane. J Biol Chem 252:5455–5467Google Scholar
  32. Stanley NR, Findlay K, Berks BC, Palmer T (2001) Escherichia coli strains blocked in Tat-dependent protein export exhibit pleiotropic defects in the cell envelope. J Bacteriol 183:139–144PubMedCrossRefGoogle Scholar
  33. Stan-Lotter H, Gupta M, Sanderson KE (1979) The influence of cations on the permeability of the outer membrane of Salmonella typhimurium and other gram-negative bacteria. Can J Microbiol 25:475–485PubMedCrossRefGoogle Scholar
  34. Treptow NA, Shuman HA (1985) Genetic evidence for substrate and periplasmic-binding-protein recognition by the MalF and MalG proteins, cytoplasmic membrane components of the Escherichia coli maltose transport system. J Bacteriol 163:654–660PubMedGoogle Scholar
  35. Tullman-Ercek D, DeLisa M, Kawarasaki Y, Iranpour P, Ribnicky B, Palmer T, Georgiou G (2007) Export pathway selectivity of Escherichia coli twin-arginine translocation signal peptides. J Biol Chem 282:8309–8316PubMedCrossRefGoogle Scholar
  36. Weiner J, Bilous PT, Shaw GM, Lubitz SP, Frost L, Thomas GH, Cole JA, Turner RJ (1998) A novel and ubiquitous system for membrane targeting and secretion of cofactor-containing proteins. Cell 93:93–101PubMedCrossRefGoogle Scholar
  37. Wexler M, Sargent F, Jack RL, Stanley NR, Bogsch EG, Robinson C, Berks BC, Palmer T (2000) TatD is a cytoplasmic protein with DNase activity. No requirement for TatD family proteins in Sec-independent protein export. J Biol Chem 275:16717–16722PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Isabelle Caldelari
    • 1
    • 2
  • Tracy Palmer
    • 1
  • Frank Sargent
    • 1
  1. 1.Division of Molecular and Environmental Microbiology, College of Life SciencesUniversity of DundeeDundeeScotland, UK
  2. 2.Institut de Biologie Moléculaire et Cellulaire du CNRS, UPR922StrasbourgFrance

Personalised recommendations