Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1993 Jul;175(14):4335–4344. doi: 10.1128/jb.175.14.4335-4344.1993

Inversion-independent phase variation of type 1 fimbriae in Escherichia coli.

M S McClain 1, I C Blomfield 1, K J Eberhardt 1, B I Eisenstein 1
PMCID: PMC204873  PMID: 8101185

Abstract

The roles of fimB and fimE in the phase-variable expression of type 1 fimbriae in Escherichia coli were examined. A method was developed to study the effects of fimB and fimE on both recombination of the fim invertible element and fimbrial expression. The method used an allelic exchange procedure consisting of two steps. The first step, construction of intermediate strains, deleted fimB and fimE. This step locked the invertible element in either the on or the off orientation. The second step of the exchange procedure introduced either wild-type or mutant alleles of fimB and/or fimE into the chromosome of the intermediate strains. Analysis of the resulting strains supported the current, plasmid-based model of recombination. Unexpectedly, strains in which the invertible element was locked in the on orientation (either by mutation of both fimB and fimE or, in a control strain, by mutation of the left inverted repeat sequence of the invertible element) continued to exhibit phase-variable expression of type 1 fimbriae. A strain in which fimA was transcribed from the tac promoter continued to exhibit phase-variable fimbrial expression, suggesting that inversion-independent phase variation cannot be explained by variable transcription initiation of fimA.

Full text

PDF
4335

Images in this article

Selected References

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

  1. Abraham J. M., Freitag C. S., Clements J. R., Eisenstein B. I. An invertible element of DNA controls phase variation of type 1 fimbriae of Escherichia coli. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5724–5727. doi: 10.1073/pnas.82.17.5724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abremski K., Gottesman S. Purification of the bacteriophage lambda xis gene product required for lambda excisive recombination. J Biol Chem. 1982 Aug 25;257(16):9658–9662. [PubMed] [Google Scholar]
  3. Adler H. I., Fisher W. D., Cohen A., Hardigree A. A. MINIATURE escherichia coli CELLS DEFICIENT IN DNA. Proc Natl Acad Sci U S A. 1967 Feb;57(2):321–326. doi: 10.1073/pnas.57.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blomfield I. C., Calie P. J., Eberhardt K. J., McClain M. S., Eisenstein B. I. Lrp stimulates phase variation of type 1 fimbriation in Escherichia coli K-12. J Bacteriol. 1993 Jan;175(1):27–36. doi: 10.1128/jb.175.1.27-36.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blomfield I. C., McClain M. S., Eisenstein B. I. Type 1 fimbriae mutants of Escherichia coli K12: characterization of recognized afimbriate strains and construction of new fim deletion mutants. Mol Microbiol. 1991 Jun;5(6):1439–1445. doi: 10.1111/j.1365-2958.1991.tb00790.x. [DOI] [PubMed] [Google Scholar]
  6. Blomfield I. C., McClain M. S., Princ J. A., Calie P. J., Eisenstein B. I. Type 1 fimbriation and fimE mutants of Escherichia coli K-12. J Bacteriol. 1991 Sep;173(17):5298–5307. doi: 10.1128/jb.173.17.5298-5307.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Blomfield I. C., Vaughn V., Rest R. F., Eisenstein B. I. Allelic exchange in Escherichia coli using the Bacillus subtilis sacB gene and a temperature-sensitive pSC101 replicon. Mol Microbiol. 1991 Jun;5(6):1447–1457. doi: 10.1111/j.1365-2958.1991.tb00791.x. [DOI] [PubMed] [Google Scholar]
  8. Blyn L. B., Braaten B. A., Low D. A. Regulation of pap pilin phase variation by a mechanism involving differential dam methylation states. EMBO J. 1990 Dec;9(12):4045–4054. doi: 10.1002/j.1460-2075.1990.tb07626.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Christie G. E., Farnham P. J., Platt T. Synthetic sites for transcription termination and a functional comparison with tryptophan operon termination sites in vitro. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4180–4184. doi: 10.1073/pnas.78.7.4180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Clark-Curtiss J. E., Curtiss R., 3rd Analysis of recombinant DNA using Escherichia coli minicells. Methods Enzymol. 1983;101:347–362. doi: 10.1016/0076-6879(83)01026-5. [DOI] [PubMed] [Google Scholar]
  11. Eisenstein B. I., Clements J. R., Dodd D. C. Isolation and characterization of a monoclonal antibody directed against type 1 fimbriae organelles from Escherichia coli. Infect Immun. 1983 Oct;42(1):333–340. doi: 10.1128/iai.42.1.333-340.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Eisenstein B. I., Dodd D. C. Pseudocatabolite repression of type 1 fimbriae of Escherichia coli. J Bacteriol. 1982 Sep;151(3):1560–1567. doi: 10.1128/jb.151.3.1560-1567.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eisenstein B. I. Phase variation of type 1 fimbriae in Escherichia coli is under transcriptional control. Science. 1981 Oct 16;214(4518):337–339. doi: 10.1126/science.6116279. [DOI] [PubMed] [Google Scholar]
  14. Freitag C. S., Abraham J. M., Clements J. R., Eisenstein B. I. Genetic analysis of the phase variation control of expression of type 1 fimbriae in Escherichia coli. J Bacteriol. 1985 May;162(2):668–675. doi: 10.1128/jb.162.2.668-675.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gerlach G. F., Clegg S. Characterization of two genes encoding antigenically distinct type-1 fimbriae of Klebsiella pneumoniae. Gene. 1988 Apr 29;64(2):231–240. doi: 10.1016/0378-1119(88)90338-1. [DOI] [PubMed] [Google Scholar]
  16. Guyer M. S., Reed R. R., Steitz J. A., Low K. B. Identification of a sex-factor-affinity site in E. coli as gamma delta. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):135–140. doi: 10.1101/sqb.1981.045.01.022. [DOI] [PubMed] [Google Scholar]
  17. Harris S. L., Elliott D. A., Blake M. C., Must L. M., Messenger M., Orndorff P. E. Isolation and characterization of mutants with lesions affecting pellicle formation and erythrocyte agglutination by type 1 piliated Escherichia coli. J Bacteriol. 1990 Nov;172(11):6411–6418. doi: 10.1128/jb.172.11.6411-6418.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Horton R. M., Hunt H. D., Ho S. N., Pullen J. K., Pease L. R. Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene. 1989 Apr 15;77(1):61–68. doi: 10.1016/0378-1119(89)90359-4. [DOI] [PubMed] [Google Scholar]
  19. Jonsson A. B., Nyberg G., Normark S. Phase variation of gonococcal pili by frameshift mutation in pilC, a novel gene for pilus assembly. EMBO J. 1991 Feb;10(2):477–488. doi: 10.1002/j.1460-2075.1991.tb07970.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Klemm P. Two regulatory fim genes, fimB and fimE, control the phase variation of type 1 fimbriae in Escherichia coli. EMBO J. 1986 Jun;5(6):1389–1393. doi: 10.1002/j.1460-2075.1986.tb04372.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kuriyama S. M., Silverblatt F. J. Effect of Tamm-Horsfall urinary glycoprotein on phagocytosis and killing of type I-fimbriated Escherichia coli. Infect Immun. 1986 Jan;51(1):193–198. doi: 10.1128/iai.51.1.193-198.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  24. McClain M. S., Blomfield I. C., Eisenstein B. I. Roles of fimB and fimE in site-specific DNA inversion associated with phase variation of type 1 fimbriae in Escherichia coli. J Bacteriol. 1991 Sep;173(17):5308–5314. doi: 10.1128/jb.173.17.5308-5314.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Murphy G. L., Connell T. D., Barritt D. S., Koomey M., Cannon J. G. Phase variation of gonococcal protein II: regulation of gene expression by slipped-strand mispairing of a repetitive DNA sequence. Cell. 1989 Feb 24;56(4):539–547. doi: 10.1016/0092-8674(89)90577-1. [DOI] [PubMed] [Google Scholar]
  26. Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ofek I., Sharon N. Lectinophagocytosis: a molecular mechanism of recognition between cell surface sugars and lectins in the phagocytosis of bacteria. Infect Immun. 1988 Mar;56(3):539–547. doi: 10.1128/iai.56.3.539-547.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ofek I., Zafriri D., Goldhar J., Eisenstein B. I. Inability of toxin inhibitors to neutralize enhanced toxicity caused by bacteria adherent to tissue culture cells. Infect Immun. 1990 Nov;58(11):3737–3742. doi: 10.1128/iai.58.11.3737-3742.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Old D. C., Duguid J. P. Selective outgrowth of fimbriate bacteria in static liquid medium. J Bacteriol. 1970 Aug;103(2):447–456. doi: 10.1128/jb.103.2.447-456.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Orndorff P. E., Falkow S. Identification and characterization of a gene product that regulates type 1 piliation in Escherichia coli. J Bacteriol. 1984 Oct;160(1):61–66. doi: 10.1128/jb.160.1.61-66.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Orndorff P. E., Spears P. A., Schauer D., Falkow S. Two modes of control of pilA, the gene encoding type 1 pilin in Escherichia coli. J Bacteriol. 1985 Oct;164(1):321–330. doi: 10.1128/jb.164.1.321-330.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ou J. T., Baron L. S., Rubin F. A., Kopecko D. J. Specific insertion and deletion of insertion sequence 1-like DNA element causes the reversible expression of the virulence capsular antigen Vi of Citrobacter freundii in Escherichia coli. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4402–4405. doi: 10.1073/pnas.85.12.4402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Pallesen L., Madsen O., Klemm P. Regulation of the phase switch controlling expression of type 1 fimbriae in Escherichia coli. Mol Microbiol. 1989 Jul;3(7):925–931. doi: 10.1111/j.1365-2958.1989.tb00242.x. [DOI] [PubMed] [Google Scholar]
  34. Purcell B. K., Pruckler J., Clegg S. Nucleotide sequences of the genes encoding type 1 fimbrial subunits of Klebsiella pneumoniae and Salmonella typhimurium. J Bacteriol. 1987 Dec;169(12):5831–5834. doi: 10.1128/jb.169.12.5831-5834.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Segal E., Billyard E., So M., Storzbach S., Meyer T. F. Role of chromosomal rearrangement in N. gonorrhoeae pilus phase variation. Cell. 1985 Feb;40(2):293–300. doi: 10.1016/0092-8674(85)90143-6. [DOI] [PubMed] [Google Scholar]
  36. Szabo M., Maskell D., Butler P., Love J., Moxon R. Use of chromosomal gene fusions to investigate the role of repetitive DNA in regulation of genes involved in lipopolysaccharide biosynthesis in Haemophilus influenzae. J Bacteriol. 1992 Nov;174(22):7245–7252. doi: 10.1128/jb.174.22.7245-7252.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weiser J. N., Maskell D. J., Butler P. D., Lindberg A. A., Moxon E. R. Characterization of repetitive sequences controlling phase variation of Haemophilus influenzae lipopolysaccharide. J Bacteriol. 1990 Jun;172(6):3304–3309. doi: 10.1128/jb.172.6.3304-3309.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zafriri D., Oron Y., Eisenstein B. I., Ofek I. Growth advantage and enhanced toxicity of Escherichia coli adherent to tissue culture cells due to restricted diffusion of products secreted by the cells. J Clin Invest. 1987 Apr;79(4):1210–1216. doi: 10.1172/JCI112939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. de Boer H. A., Comstock L. J., Vasser M. The tac promoter: a functional hybrid derived from the trp and lac promoters. Proc Natl Acad Sci U S A. 1983 Jan;80(1):21–25. doi: 10.1073/pnas.80.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES