Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Jul;169(7):3001–3006. doi: 10.1128/jb.169.7.3001-3006.1987

Mu d-directed lacZ fusions regulated by low pH in Escherichia coli.

J L Slonczewski, T N Gonzalez, F M Bartholomew, N J Holt
PMCID: PMC212340  PMID: 2954947

Abstract

Methods were devised to isolate strains of Escherichia coli containing Mu d (lacZ Kmr) operon fusions regulated by external pH and by internal pH. External acid-inducible fusions (exa) were detected by plating a Mu d fusion pool on Luria broth with 5-bromo-4-chloro-3-indolyl-beta-D-galactoside, buffered at pH 7.4, and then replica plating on the same medium buffered at pH 5.5. Two exa strains showed induction by external acidification, up to 800-fold and 90-fold. Induction of both fusions was maximal at pH 5.6 and minimal over pH 7.0 to 8.3. There was no induction by membrane-permeable weak acids which depress internal pH at constant external pH. Anaerobiosis increased the steady-state level of transcription of exa-1 5-fold and of exa-2 2.5-fold at low external pH. Internal acid-inducible fusions (ina) were detected by plating a Mu d fusion pool on MacConkey medium, pH 6.8, and then replica plating with 15 mM benzoate. Two ina strains showed 10-fold induction by 20 mM benzoate at external pH 7.0. Similar results were obtained with other weak acids; their relative potency (salicylate greater than benzoate greater than dimethoxazoledinedione) was consistent with their relative ability to depress internal pH. In the absence of a weak acid, external pH had almost no effect over the pH range 5.5 to 8.0. Anaerobiosis did not affect ina induction. To our knowledge, this is the first report of E. coli genes induced specifically by internal but not external acidification and the first report of gene fusions induced by external acidification but not by weak acids.

Full text

PDF
3001

Selected References

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

  1. Booth I. R. Regulation of cytoplasmic pH in bacteria. Microbiol Rev. 1985 Dec;49(4):359–378. doi: 10.1128/mr.49.4.359-378.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Casadaban M. J., Cohen S. N. 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 U S A. 1979 Sep;76(9):4530–4533. doi: 10.1073/pnas.76.9.4530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Castilho B. A., Olfson P., Casadaban M. J. Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons. J Bacteriol. 1984 May;158(2):488–495. doi: 10.1128/jb.158.2.488-495.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ferguson W. J., Braunschweiger K. I., Braunschweiger W. R., Smith J. R., McCormick J. J., Wasmann C. C., Jarvis N. P., Bell D. H., Good N. E. Hydrogen ion buffers for biological research. Anal Biochem. 1980 May 15;104(2):300–310. doi: 10.1016/0003-2697(80)90079-2. [DOI] [PubMed] [Google Scholar]
  5. Gale E. F., Epps H. M. The effect of the pH of the medium during growth on the enzymic activities of bacteria (Escherichia coli and Micrococcus lysodeikticus) and the biological significance of the changes produced. Biochem J. 1942 Sep;36(7-9):600–618. doi: 10.1042/bj0360600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Good N. E., Winget G. D., Winter W., Connolly T. N., Izawa S., Singh R. M. Hydrogen ion buffers for biological research. Biochemistry. 1966 Feb;5(2):467–477. doi: 10.1021/bi00866a011. [DOI] [PubMed] [Google Scholar]
  7. Gottesman S. Bacterial regulation: global regulatory networks. Annu Rev Genet. 1984;18:415–441. doi: 10.1146/annurev.ge.18.120184.002215. [DOI] [PubMed] [Google Scholar]
  8. Hall M. N., Silhavy T. J. Genetic analysis of the major outer membrane proteins of Escherichia coli. Annu Rev Genet. 1981;15:91–142. doi: 10.1146/annurev.ge.15.120181.000515. [DOI] [PubMed] [Google Scholar]
  9. Holt R. A., Stephens G. M., Morris J. G. Production of Solvents by Clostridium acetobutylicum Cultures Maintained at Neutral pH. Appl Environ Microbiol. 1984 Dec;48(6):1166–1170. doi: 10.1128/aem.48.6.1166-1170.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kashket E. R. Stoichiometry of the H+-ATPase of Escherichia coli cells during anaerobic growth. FEBS Lett. 1983 Apr 18;154(2):343–346. doi: 10.1016/0014-5793(83)80179-3. [DOI] [PubMed] [Google Scholar]
  11. Kihara M., Macnab R. M. Cytoplasmic pH mediates pH taxis and weak-acid repellent taxis of bacteria. J Bacteriol. 1981 Mar;145(3):1209–1221. doi: 10.1128/jb.145.3.1209-1221.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kobayashi H. A proton-translocating ATPase regulates pH of the bacterial cytoplasm. J Biol Chem. 1985 Jan 10;260(1):72–76. [PubMed] [Google Scholar]
  13. Kobayashi H., Suzuki T., Kinoshita N., Unemoto T. Amplification of the Streptococcus faecalis proton-translocating ATPase by a decrease in cytoplasmic pH. J Bacteriol. 1984 Jun;158(3):1157–1160. doi: 10.1128/jb.158.3.1157-1160.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Le Rudulier D., Strom A. R., Dandekar A. M., Smith L. T., Valentine R. C. Molecular biology of osmoregulation. Science. 1984 Jun 8;224(4653):1064–1068. doi: 10.1126/science.224.4653.1064. [DOI] [PubMed] [Google Scholar]
  15. Neidhardt F. C., VanBogelen R. A., Vaughn V. The genetics and regulation of heat-shock proteins. Annu Rev Genet. 1984;18:295–329. doi: 10.1146/annurev.ge.18.120184.001455. [DOI] [PubMed] [Google Scholar]
  16. Padan E., Zilberstein D., Rottenberg H. The proton electrochemical gradient in Escherichia coli cells. Eur J Biochem. 1976 Apr 1;63(2):533–541. doi: 10.1111/j.1432-1033.1976.tb10257.x. [DOI] [PubMed] [Google Scholar]
  17. Padan E., Zilberstein D., Schuldiner S. pH homeostasis in bacteria. Biochim Biophys Acta. 1981 Dec;650(2-3):151–166. doi: 10.1016/0304-4157(81)90004-6. [DOI] [PubMed] [Google Scholar]
  18. Repaske D. R., Adler J. Change in intracellular pH of Escherichia coli mediates the chemotactic response to certain attractants and repellents. J Bacteriol. 1981 Mar;145(3):1196–1208. doi: 10.1128/jb.145.3.1196-1208.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rosner J. L. Nonheritable resistance to chloramphenicol and other antibiotics induced by salicylates and other chemotactic repellents in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8771–8774. doi: 10.1073/pnas.82.24.8771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Salmond C. V., Kroll R. G., Booth I. R. The effect of food preservatives on pH homeostasis in Escherichia coli. J Gen Microbiol. 1984 Nov;130(11):2845–2850. doi: 10.1099/00221287-130-11-2845. [DOI] [PubMed] [Google Scholar]
  21. Schuldiner S., Agmon V., Brandsma J., Cohen A., Friedman E., Padan E. Induction of SOS functions by alkaline intracellular pH in Escherichia coli. J Bacteriol. 1986 Nov;168(2):936–939. doi: 10.1128/jb.168.2.936-939.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Silhavy T. J., Beckwith J. R. Uses of lac fusions for the study of biological problems. Microbiol Rev. 1985 Dec;49(4):398–418. doi: 10.1128/mr.49.4.398-418.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Slonczewski J. L., Macnab R. M., Alger J. R., Castle A. M. Effects of pH and repellent tactic stimuli on protein methylation levels in Escherichia coli. J Bacteriol. 1982 Oct;152(1):384–399. doi: 10.1128/jb.152.1.384-399.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Slonczewski J. L., Rosen B. P., Alger J. R., Macnab R. M. pH homeostasis in Escherichia coli: measurement by 31P nuclear magnetic resonance of methylphosphonate and phosphate. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6271–6275. doi: 10.1073/pnas.78.10.6271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Taglicht D., Padan E., Oppenheim A. B., Schuldiner S. An alkaline shift induces the heat shock response in Escherichia coli. J Bacteriol. 1987 Feb;169(2):885–887. doi: 10.1128/jb.169.2.885-887.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Walker G. C. Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev. 1984 Mar;48(1):60–93. doi: 10.1128/mr.48.1.60-93.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wanner B. L., McSharry R. Phosphate-controlled gene expression in Escherichia coli K12 using Mudl-directed lacZ fusions. J Mol Biol. 1982 Jul 5;158(3):347–363. doi: 10.1016/0022-2836(82)90202-9. [DOI] [PubMed] [Google Scholar]
  28. Wanner B. L. Overlapping and separate controls on the phosphate regulon in Escherichia coli K12. J Mol Biol. 1983 May 25;166(3):283–308. doi: 10.1016/s0022-2836(83)80086-2. [DOI] [PubMed] [Google Scholar]
  29. Winkelman J. W., Clark D. P. Anaerobically induced genes of Escherichia coli. J Bacteriol. 1986 Jul;167(1):362–367. doi: 10.1128/jb.167.1.362-367.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yerkes J. H., Casson L. P., Honkanen A. K., Walker G. C. Anaerobiosis induces expression of ant, a new Escherichia coli locus with a role in anaerobic electron transport. J Bacteriol. 1984 Apr;158(1):180–186. doi: 10.1128/jb.158.1.180-186.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zilberstein D., Agmon V., Schuldiner S., Padan E. Escherichia coli intracellular pH, membrane potential, and cell growth. J Bacteriol. 1984 Apr;158(1):246–252. doi: 10.1128/jb.158.1.246-252.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zilberstein D., Agmon V., Schuldiner S., Padan E. The sodium/proton antiporter is part of the pH homeostasis mechanism in Escherichia coli. J Biol Chem. 1982 Apr 10;257(7):3687–3691. [PubMed] [Google Scholar]

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

RESOURCES