Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 Jun;173(12):3609–3614. doi: 10.1128/jb.173.12.3609-3614.1991

Leakage of periplasmic enzymes from envA1 strains of Escherichia coli.

K Young 1, L L Silver 1
PMCID: PMC207986  PMID: 1904854

Abstract

Previous work ascribed antibiotic hypersensitivity of the envA1 mutant to lowered lipopolysaccharide levels and exposure of the lipid bilayer. In the detailed characterization of the EnvA permeability phenotype presented here, the envA1 mutation was shown to confer leakage of the periplasmic enzymes beta-lactamase and RNase I. Leakage was observed in three different genetic backgrounds, including the original envA1 strain and its parent. In contrast, no detectable leakage of the cytoplasmic enzyme beta-galactosidase was observed. Sensitivity of envA1 strains to a range of antibiotics not previously reported was tested, and lipophilicity (partition coefficient) of a number of antibiotics was determined. On the basis of observations of periplasmic leakage and sensitivity to large hydrophilic antibiotics and lysozyme, part of the permeability phenotype of the envA1 mutant is proposed to be due to transient rupture and resealing of the EDTA-sensitive outer membrane layer. In this regard, the EnvA permeability phenotype falls into a general class of permeability/leaky mutants of both Escherichia coli and Salmonella typhimurium.

Full text

PDF

Images in this article

3612Image
on p.3612

Selected References

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

  1. Angus B. L., Carey A. M., Caron D. A., Kropinski A. M., Hancock R. E. Outer membrane permeability in Pseudomonas aeruginosa: comparison of a wild-type with an antibiotic-supersusceptible mutant. Antimicrob Agents Chemother. 1982 Feb;21(2):299–309. doi: 10.1128/aac.21.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ashby J., Piddock L. J., Wise R. An investigation of the hydrophobicity of the quinolones. J Antimicrob Chemother. 1985 Dec;16(6):805–808. doi: 10.1093/jac/16.6.805-a. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Beall B., Lutkenhaus J. Sequence analysis, transcriptional organization, and insertional mutagenesis of the envA gene of Escherichia coli. J Bacteriol. 1987 Dec;169(12):5408–5415. doi: 10.1128/jb.169.12.5408-5415.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  6. Boman H. G., Eriksson-Grennberg K. G., Normark S., Matsson E. Resistance of Escherichia coli to penicillins. IV. Genetic study of mutants resistant to D,L-ampicillin concentrations o 100 mu-g-ml. Genet Res. 1968 Oct;12(2):169–185. doi: 10.1017/s0016672300011782. [DOI] [PubMed] [Google Scholar]
  7. Boman H. G., Nordström K., Normark S. Penicillin resistance in Escherichia coli K12: synergism between penicillinases and a barrier in the outer part of the envelope. Ann N Y Acad Sci. 1974 May 10;235(0):569–586. doi: 10.1111/j.1749-6632.1974.tb43291.x. [DOI] [PubMed] [Google Scholar]
  8. Coleman W. G., Jr, Leive L. Two mutations which affect the barrier function of the Escherichia coli K-12 outer membrane. J Bacteriol. 1979 Sep;139(3):899–910. doi: 10.1128/jb.139.3.899-910.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fung J., MacAlister T. J., Rothfield L. I. Role of murein lipoprotein in morphogenesis of the bacterial division septum: phenotypic similarity of lkyD and lpo mutants. J Bacteriol. 1978 Mar;133(3):1467–1471. doi: 10.1128/jb.133.3.1467-1471.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grundström T., Jaurin B., Edlund T., Normark S. Physical mapping and expression of hybrid plasmids carrying chromosomal beta-lactamase genes of Escherichia coli K-12. J Bacteriol. 1980 Sep;143(3):1127–1134. doi: 10.1128/jb.143.3.1127-1134.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grundström T., Normark S., Magnusson K. E. Overproduction of outer membrane protein suppresses envA-induced hyperpermeability. J Bacteriol. 1980 Dec;144(3):884–890. doi: 10.1128/jb.144.3.884-890.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hirota Y., Suzuki H., Nishimura Y., Yasuda S. On the process of cellular division in Escherichia coli: a mutant of E. coli lacking a murein-lipoprotein. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1417–1420. doi: 10.1073/pnas.74.4.1417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Jones C., Holland I. B. Role of the SulB (FtsZ) protein in division inhibition during the SOS response in Escherichia coli: FtsZ stabilizes the inhibitor SulA in maxicells. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6045–6049. doi: 10.1073/pnas.82.18.6045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Laminet A. A., Plückthun A. The precursor of beta-lactamase: purification, properties and folding kinetics. EMBO J. 1989 May;8(5):1469–1477. doi: 10.1002/j.1460-2075.1989.tb03530.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Leive L. Release of lipopolysaccharide by EDTA treatment of E. coli. Biochem Biophys Res Commun. 1965 Nov 22;21(4):290–296. doi: 10.1016/0006-291x(65)90191-9. [DOI] [PubMed] [Google Scholar]
  18. Nakae T. Outer-membrane permeability of bacteria. Crit Rev Microbiol. 1986;13(1):1–62. doi: 10.3109/10408418609108734. [DOI] [PubMed] [Google Scholar]
  19. Nikaido H. Outer membrane of Salmonella typhimurium. Transmembrane diffusion of some hydrophobic substances. Biochim Biophys Acta. 1976 Apr 16;433(1):118–132. doi: 10.1016/0005-2736(76)90182-6. [DOI] [PubMed] [Google Scholar]
  20. Nikaido H., Vaara M. Molecular basis of bacterial outer membrane permeability. Microbiol Rev. 1985 Mar;49(1):1–32. doi: 10.1128/mr.49.1.1-32.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Normark S., Boman H. G., Bloom G. D. Cell division in a chain-forming envA mutant of Escherichia coli K12. Fine structure of division sites and effects of EDTA, lysozyme and ampicillin. Acta Pathol Microbiol Scand B Microbiol Immunol. 1971;79(5):651–664. doi: 10.1111/j.1699-0463.1971.tb00093.x. [DOI] [PubMed] [Google Scholar]
  22. Normark S., Boman H. G., Matsson E. Mutant of Escherichia coli with anomalous cell division and ability to decrease episomally and chromosomally mediated resistance to ampicillin and several other antibiotics. J Bacteriol. 1969 Mar;97(3):1334–1342. doi: 10.1128/jb.97.3.1334-1342.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Normark S., Burman L. G. Resistance of Escherichia coli to penicillins: fine-structure mapping and dominance of chromosomal beta-lactamase mutations. J Bacteriol. 1977 Oct;132(1):1–7. doi: 10.1128/jb.132.1.1-7.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Normark S. Genetics of a chain-forming mutant of Escherichia coli. Transduction and dominance of the envA gene mediating increased penetration to some antibacterial agents. Genet Res. 1970 Aug;16(1):63–78. doi: 10.1017/s0016672300002287. [DOI] [PubMed] [Google Scholar]
  25. Normark S., Wolf-Watz H. Cell division and permeability of unbalanced envelope mutants of Escherichia coli K12. Ann Microbiol (Paris) 1974 Sep;125 B(2):211–226. [PubMed] [Google Scholar]
  26. O'Callaghan C. H., Morris A., Kirby S. M., Shingler A. H. Novel method for detection of beta-lactamases by using a chromogenic cephalosporin substrate. Antimicrob Agents Chemother. 1972 Apr;1(4):283–288. doi: 10.1128/aac.1.4.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Roantree R. J., Kuo T. T., MacPhee D. G. The effect of defined lipopolysaccharide core defects upon antibiotic resistances of Salmonella typhimurium. J Gen Microbiol. 1977 Dec;103(2):223–234. doi: 10.1099/00221287-103-2-223. [DOI] [PubMed] [Google Scholar]
  28. Scudamore R. A., Beveridge T. J., Goldner M. Outer-membrane penetration barriers as components of intrinsic resistance to beta-lactam and other antibiotics in Escherichia coli K-12. Antimicrob Agents Chemother. 1979 Feb;15(2):182–189. doi: 10.1128/aac.15.2.182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sigal I. S., DeGrado W. F., Thomas B. J., Petteway S. R., Jr Purification and properties of thiol beta-lactamase. A mutant of pBR322 beta-lactamase in which the active site serine has been replaced with cysteine. J Biol Chem. 1984 Apr 25;259(8):5327–5332. [PubMed] [Google Scholar]
  30. Vaara M. Antimicrobial susceptibility of Salmonella typhimurium carrying the outer membrane permeability mutation SS-B. Antimicrob Agents Chemother. 1990 May;34(5):853–857. doi: 10.1128/aac.34.5.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weigand R. A., Rothfield L. I. Genetic and physiological classification of periplasmic-leaky mutants of Salmonella typhimurium. J Bacteriol. 1976 Jan;125(1):340–345. doi: 10.1128/jb.125.1.340-345.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weigand R. A., Vinci K. D., Rothfield L. I. Morphogenesis of the bacterial division septum: a new class of septation-defective mutants. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1882–1886. doi: 10.1073/pnas.73.6.1882. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wijsman H. J., Koopman C. R. The relation of the genes envA and ftsA in Escherichia coli. Mol Gen Genet. 1976 Aug 10;147(1):99–102. doi: 10.1007/BF00337942. [DOI] [PubMed] [Google Scholar]
  34. Wolf-Watz H., Normark S., Bloom G. D. Rapid method for isolation of large quantities of outer membrane from Escherichia coli K-12 and its application to the study of envelope mutants. J Bacteriol. 1973 Sep;115(3):1191–1197. doi: 10.1128/jb.115.3.1191-1197.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES