Skip to main content
SpringerLink
Log in

Transient, specific and extremely rapid release of osmolytes from growing cells of Escherichia coli K-12 exposed to hypoosmotic shock

  • Original Papers
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

The influence of hypoosmotic shock on the solute content of growing Escherichia coli K-12 cells was investigated at 37°C. Within 20 s after the shock the cells had released most of their osmolytes K+, glutamate and trehalose. This release was specific and not due to rupture of the cell membrane, since under these conditions i) the cells neither lost protein nor ATP, ii)[14C]-labeled sucrose did not enter the cytoplasm from the periplasm, and iii) except for their glutamate and aspartate level, which decreased, the amino acid pool of alanine, lysine and arginine of the cells remained approximately constant. Within a minute after the shock the cells started to reaccumulate parts of their previously released glutamate, aspartate and K+, but not trehalose and resumed growth within 10 min after the shock. Experiments with K+-transport mutants showed that none of the genetically-identified K+ transport systems is involved in the K+-release process. Reaccumulation of K+ took place via the uptake systems TrkG and TrkH. The possibility is discussed that the exit of solutes after hypoosmotic shock occurs via several stretch-activated channels, which each allow the release of a specific osmolyte.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

OD578 :

optical density at 578 nm

TEA:

triethylammonium

TMG:

1,-S-methyl-β-thiogalactopyranoside

References

  • Anraku Y (1978) Active transport of amino acids. In: Rosen BP (ed) Bacterial transport. Marcel Dekker, New York, pp 171–219.

    Google Scholar 

  • Bakker EP (1992a) Cell K+ and K+ transport systems in prokaryotes. In: Bakker EP (ed) Alkali cation transport systems in prokaryotes. CRC Press, Boca Raton, Florida, pp 205–224.

    Google Scholar 

  • Bakker EP (1992b) Low affinity K+ uptake systems. In: Bakker EP (ed) Alkali cation transport systems in prokaryotes. CRC Press, Boca Raton, Florida, pp 253–276.

    Google Scholar 

  • Bakker EP, Mangerich WE (1981) Interconversion of components of the bacterial proton-motive force by electrogenic potassium transport. J Bacteriol 147: 820–826.

    Google Scholar 

  • Bakker EP, Booth IR, Dinnbier U, Epstein W, Gajewska A (1987) Evidence for multiple K+ export systems in Escherichia coli. J Bacteriol 169: 3743–3749.

    Google Scholar 

  • Benz R, Bauer K (1988) Permeation of hydrophilic molecules through the outer membrane of gram-negative bacteria. Review on bacterial porins. Eur J Biochem 176: 1–19.

    Google Scholar 

  • Berrier C, Coulombe A, Houssin C, Ghazi A (1989) A patch-clamp study of ion channels of inner and outer membranes and of contact zones of Escherichia coli. FEBS Lett 259: 27–32.

    Google Scholar 

  • Berrier G, Coulombe A, Szabo I, Zoratti M, Ghazi A (1992) Gadolinium ion inhibits loss of metabolites induced by osmotic shock and large stretch activated channels in bacteria. Eur J Biochem 206: 559–565.

    Google Scholar 

  • Boos W, Ehmann U, Bremer E, Middendorf A, Postma P (1987) Trehalase of Escherichia coli. Mapping and cloning of its structural gene and identification of the enzyme as a periplasmic protein induced under high osmolarity growth conditions. J Biol Chem 262: 3450–3461.

    Google Scholar 

  • Booth IR (1985) Regulation of cytoplasmic pH in bacteria. Microbiol Rev 49: 359–378.

    Google Scholar 

  • Booth IR (1992) Regulation of gene expression during osmoregulation: the role of potassium as a secondary signal of osmotic stress. In: Bakker EP (ed) Alkali cation transport systems in prokaryotes. CRC Press, Boca Raton, Florida, pp 309–331.

    Google Scholar 

  • Booth IR, Higgins CF (1990) Enteric bacteria and osmotic stress: intracellular potassium glutamate as a secondary signal of osmotic stress? FEMS Microbiol Rev 75: 239–246.

    Google Scholar 

  • Bossemeyer D, Borchard A, Dosch DC, Helmer GC, Epstein W, Booth IR, Bakker EP (1989) K+-transport protein TrkA of Escherichia coli is a peripheral membrane protein that requires other trk-gene products for attachment to the cytoplasmic membrane. J Biol Chem 264: 16403–16410.

    Google Scholar 

  • Bremer E, Silhavy T, Weiseman J, Weinstock G (1984) λplacMu: a transposable derivative of bacteriophage lambda for creating lacZ fusions in a single step. J Bacteriol 158: 1084–1093.

    Google Scholar 

  • Bremer E, Silhavy T, Weinstock G (1985) Transposable λplacMu bacteriophages for creating lacZ operon fusions and kanamycin resistance insertions in Escherichia coli. J Bacteriol 162: 1092–1099.

    Google Scholar 

  • Cairney J, Booth IR, Higgins CF (1985a) Salmonella typhimurium proP gene encodes a transport system for the osmoprotectant betaine. J Bacteriol 164: 1218–1223.

    Google Scholar 

  • Cairney J, Booth IR, Higgins CF (1985b) Osmoregulation of gene expression in Salmonella typhimurium: proU encodes an osmotically induced betaine transport system. J Bacteriol 164: 1224–1232.

    Google Scholar 

  • Csonka LN (1989) Physiological and genetic responses of bacteria to osmotic stress. Microbiol Rev 53: 121–147.

    Google Scholar 

  • Dinnbier U, Limpinsel E, Schmid R, Bakker EP (1988) Transient accumulation of potassium glutamate and its replacement by trchalose during adaptation of growing cells of Escherichia coli K-12 to elevated sodium chloride concentrations. Arch Microbiol 150: 348–357.

    Google Scholar 

  • Dosch DC, Helmer GL, Sutton FF, Salvacion FF, Epstein W (1991) Genetic analysis of potassium transport loci in Escherichia coli: evidence for three constitutive systems mediating uptake of potassium. J Bacteriol 173: 687–696.

    Google Scholar 

  • Epstein W (1986) Osmoregulation by potassium transport in Escherichia coli. FEMS Microbiol Rev 39: 73–78.

    Google Scholar 

  • Epstein W, Kim BS (1971) Potassium transport in Escherichia coli K-12. J Bacteriol 108: 639–644.

    Google Scholar 

  • Garlid KD, DiResta DJ, Beavis AD, Martin WH (1986) On the mechanism by which dicyclohexylcarbodiimide and quinine inhibit K+ transport in rat liver mitochondria. J Biol Chem 261: 1529–1535.

    Google Scholar 

  • Grothe S, Krogsrud RL, McLellan DJ, Milner JL, Wood JM (1986) Proline transport and osmotic stress response in Escherichia coli K-12. J Bacteriol 166: 253–259.

    Google Scholar 

  • Hanson RS, Philips JA (1981) Chemical composition. In: Gerhardt P, Murray RGE, Costilow RN, Nester EW, Wood WA, Krieg NR, Phillips GB (eds) Manual on methods for general bacteriology. American Society for Microbiology, Washington, DC, pp 328–364.

    Google Scholar 

  • Hoischen C, Krämer R (1989) Evidence for an efflux carrier system involved in the secretion of glutamate by Corynebacterium glutamicum. Arch Microbiol 151: 342–347.

    Google Scholar 

  • Jebbar M, Talibart R, Gloux K, Bernard T, Blanco C (1992) Osmoprotection of Escherichia coli by ectoine: uptake and accumulation characteristics. J Bacteriol 174: 5027–5035.

    Google Scholar 

  • Kashket ER, Barker SL (1977) Effect of potassium on the electrical and pH gradient across the membrane of Streptococcus lactis. J Bacteriol 130: 1017–1023.

    Google Scholar 

  • Kimmich GA, Randles J, Brand JS (1975) Assay of picomole amounts of ATP, ADP and AMP using the luciferine enzyme system. Anal Biochem 69: 187–206.

    Google Scholar 

  • Konings WN, Poolman B, Driessen A (1992) Can the excretion of metabolites by bacteria be manipulated? FEMS Microbiol Rev 93: 93–108.

    Google Scholar 

  • Koo S-P, Higgins CF, Booth IR (1991) Regulation of compatible solute accumulation in Salmonella typhimurium: evidence for a glycine betaine efflux system. J Gen Microbiol 137: 2617–2625.

    Google Scholar 

  • Larsen PI, Sydnes LK, Landfald B, Strøm AR (1987) Osmoregulation in Escherichia coli by accumulation of organic osmolytes: betaines, glutamic acid, and trehalose. Arch Microbiol 147: 1–7.

    Google Scholar 

  • Martinac B, Buechner M, Delcour AH, Adler J, Kung C (1987) Pressure-sensitive ion channels in Escherichia coli. Proc Natl Acad Sci USA 84: 2297–2301.

    Google Scholar 

  • McLaggan D, Epstein W (1991) Escherichia coli accumulates the eukaryotic osmolyte taurine at high osmolarity. FEMS Microbiol Lett 81: 209–214.

    Google Scholar 

  • Measures JC (1975) Role of amino acids in osmoregulation of non-halophilic bacteria. Nature 257: 398–400.

    Google Scholar 

  • Meury J, Robin A, Monnier-Champeix P (1985) Turgor-controlled K+ fluxes and their pathways in Escherichia coli. Eur J Biochem 151: 613–619.

    Google Scholar 

  • Michels M, Bakker EP (1987) Low affinity potassium uptake system in Bacillus acidocaldarius. J Bacteriol 169: 4335–4341.

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  • Morris C (1990) Mechanosensitive ion channels. J Membrane Biol 113: 93–107.

    Google Scholar 

  • Ohwada T, Sagisaka S (1987) An immediate and steep increase in ATP concentration in response to reduced turgor pressure in Escherichia coli B. Arch Biochem Biophys 259: 157–163.

    Google Scholar 

  • Ohwada T, Sagisaka S (1988) The differential roles of potassium, proline, and betaine in osmoregulation of Escherichia coli B. Agric Biol Chem 52: 313–320.

    Google Scholar 

  • Padan E, Schuldiner S (1992) Na+ transport systems in prokaryotes. In: Bakker EP (ed) Alkali cation transport systems in prokaryotes. CRC Press, Boca Raton, Florida, pp 3–24.

    Google Scholar 

  • Rhoads DB, Epstein W (1977) Energy coupling to net K+ transport in Escherichia coli K-12. J Biol Chem 252: 1394–1401.

    Google Scholar 

  • Rhoads DB, Epstein W (1978) Cation transport in Escherichia coli. IX Regulation of K+ transport. J Gen Physiol 72: 283–295.

    Google Scholar 

  • Rhoads DB, Waters FB, Epstein W (1976) Cation transport in Escherichia coli. VIII. Potassium transport mutants. J Gen Physiol 67: 325–341.

    Google Scholar 

  • Richey B, Cayley DS, Mossing MC, Kolka C, Anderson CF, Farrar TC, RecordJr MT (1987) Variability of the intracellular ionic environment of Escherichia coli. Differences between in vitro and in vivo effects of ion concentration on protein-DNA interactions and gene expression. J Biol Chem 262: 7157–7164.

    Google Scholar 

  • Schlösser A, Hamann A, Bossemeyer D, Schneider E, Bakker EP (1993) NAD+ binding to the Escherichia coli K+-uptake protein TrkA and sequence similarity between TrkA and domains of a family of dehydrogenases suggest a role for NAD+ in bacterial transport. Mol Microbiol (in press).

  • Shiio I, Otsuka SI, Takahashi M (1962) Effect of biotin on the bacterial formation of glutamic acid. I. Glutamate formation and cellular permeability of amino acids. J Biochem 51: 56–62.

    Google Scholar 

  • Silver S, Walderhaug M (1992) Gene regulation of plasmid- and chromosome-determined inorganic ion transport in bacteria. Microbiol Rev 56: 195–228.

    Google Scholar 

  • Stock JB, Rauch B, Roseman S (1977) Periplasmic space in Salmonella typhimurium and Escherichia coli. J Biol Chem 252: 7850–7861.

    Google Scholar 

  • Strøm AR, Falkenberg P, Landfald B (1986) Genetics of osmoregulation in Escherichia coli: uptake and biosynthesis of organic osmolytes. FEMS Microbiol Rev 39: 79–86.

    Google Scholar 

  • Styrvold OB, Strøm AR (1991) Synthesis, accumulation, and excretion of trehalose in osmotically stressed Escherichia coli K-12 strains: influence of amber suppressors and function of the periplasmic trehalase. J Bacteriol 173: 1187–1192.

    Google Scholar 

  • Sutherland L, Cairney J, Elmore MJ, Booth IR, Higgins CF (1987) Osmotic regulation of transcription: induction of the proU betaine transport gene is dependent on accumulation of intracellular potassium. J Bacteriol 168: 805–814.

    Google Scholar 

  • Tempest DW, Meers JJ, Brown CM (1970) Influence of environment on the content and composition of microbial free amino acid pools. J Gen Microbiol 64: 171–185.

    Google Scholar 

  • Tsapis A, Kepes A (1977) Transient breakdown of the permeability barrier of the membrane of Escherichia coli upon hypoosmotic shock. Biochim Biophys Acta 469: 1–12.

    Google Scholar 

  • Tschichholz I, Trüper HG (1990) Fate of compatible solutes during dilution stress in Ectothiorhodospira halochloris. FEMS Microbiol Ecol 73: 181–186.

    Google Scholar 

  • Walderhaug MO, Dosch DC, Epstein W (1987) Potassium transport in bacteria. In: Rosen BP, Silver S (ed) Ion transport in bacteria. Academic Press, New York, pp 84–130.

    Google Scholar 

  • Wiggins P (1990) Role of water in some biological processes. Microbiol Rev 54: 432–449.

    Google Scholar 

  • Zoratti M, Ghazi A (1992) Stretch-activated channels in prokaryotes. In: Bakker EP (ed) Alkali cation transport systems in prokaryotes. CRC Press, Boca Raton, Florida, pp 349–358.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Fachbereich Mikrobiologie, Universität Osnabrück, Postfach 4469, D-49069, Osnabrück, Germany

    Manfred Schleyer, Roland Schmid & Evert P. Bakker

Authors
  1. Manfred Schleyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Roland Schmid
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Evert P. Bakker
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schleyer, M., Schmid, R. & Bakker, E.P. Transient, specific and extremely rapid release of osmolytes from growing cells of Escherichia coli K-12 exposed to hypoosmotic shock. Arch. Microbiol. 160, 424–431 (1993). https://doi.org/10.1007/BF00245302

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00245302

Key words

Search

Navigation