Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1993 May;175(9):2770–2778. doi: 10.1128/jb.175.9.2770-2778.1993

Rapid, synchronous, and stable induction of type 1 piliation in Escherichia coli by using a chromosomal lacUV5 promoter.

L D Woodall 1, P W Russell 1, S L Harris 1, P E Orndorff 1
PMCID: PMC204584  PMID: 8097517

Abstract

Type 1 pili are filamentous proteinaceous appendages produced by certain members of the family Enterobacteriaceae. In Escherichia coli, the adhesive properties of these pili are due to the binding of at least one minor pilus component to mannose, a sugar common to cell surface molecules of many eukaryotic cells. The study of pilus assembly may be benefited by a rapid way of inducing pilus synthesis de novo. We describe herein the construction and characterization of a strain in which piliation can be rapidly induced by the addition of lactose or its analog isopropyl-beta-D-thiogalactopyranoside. This was accomplished by placing the chromosomal fimA gene (encoding the major structural subunit of pili) under lacUV5 promoter control. Further experiments suggested that transcription of genes downstream of fimA, whose products are required for normal pilus assembly and function, may also be controlled by the lacUV5 promoter. The construction described herein may have a variety of applications apart from aiding the study of pilus assembly since its adhesive properties can be rapidly and easily turned on and off.

Full text

PDF
2770

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. Auerswald E. A., Ludwig G., Schaller H. Structural analysis of Tn5. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):107–113. doi: 10.1101/sqb.1981.045.01.019. [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. Backman K., Ptashne M., Gilbert W. Construction of plasmids carrying the cI gene of bacteriophage lambda. Proc Natl Acad Sci U S A. 1976 Nov;73(11):4174–4178. doi: 10.1073/pnas.73.11.4174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bloch C. A., Orndorff P. E. Impaired colonization by and full invasiveness of Escherichia coli K1 bearing a site-directed mutation in the type 1 pilin gene. Infect Immun. 1990 Jan;58(1):275–278. doi: 10.1128/iai.58.1.275-278.1990. [DOI] [PMC free article] [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. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Göransson M., Sondén B., Nilsson P., Dagberg B., Forsman K., Emanuelsson K., Uhlin B. E. Transcriptional silencing and thermoregulation of gene expression in Escherichia coli. Nature. 1990 Apr 12;344(6267):682–685. doi: 10.1038/344682a0. [DOI] [PubMed] [Google Scholar]
  9. Hanson M. S., Brinton C. C., Jr Identification and characterization of E. coli type-1 pilus tip adhesion protein. Nature. 1988 Mar 17;332(6161):265–268. doi: 10.1038/332265a0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Jorgensen R. A., Berg D. E., Allet B., Reznikoff W. S. Restriction enzyme cleavage map of Tn10, a transposon which encodes tetracycline resistance. J Bacteriol. 1979 Jan;137(1):681–685. doi: 10.1128/jb.137.1.681-685.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kawula T. H., Orndorff P. E. Rapid site-specific DNA inversion in Escherichia coli mutants lacking the histonelike protein H-NS. J Bacteriol. 1991 Jul;173(13):4116–4123. doi: 10.1128/jb.173.13.4116-4123.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Klemm P., Christiansen G. The fimD gene required for cell surface localization of Escherichia coli type 1 fimbriae. Mol Gen Genet. 1990 Jan;220(2):334–338. doi: 10.1007/BF00260505. [DOI] [PubMed] [Google Scholar]
  14. Klemm P., Krogfelt K. A., Hedegaard L., Christiansen G. The major subunit of Escherichia coli type 1 fimbriae is not required for D-mannose-specific adhesion. Mol Microbiol. 1990 Apr;4(4):553–559. doi: 10.1111/j.1365-2958.1990.tb00623.x. [DOI] [PubMed] [Google Scholar]
  15. Krogfelt K. A., Bergmans H., Klemm P. Direct evidence that the FimH protein is the mannose-specific adhesin of Escherichia coli type 1 fimbriae. Infect Immun. 1990 Jun;58(6):1995–1998. doi: 10.1128/iai.58.6.1995-1998.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Krogfelt K. A., Klemm P. Investigation of minor components of Escherichia coli type 1 fimbriae: protein chemical and immunological aspects. Microb Pathog. 1988 Mar;4(3):231–238. doi: 10.1016/0882-4010(88)90073-3. [DOI] [PubMed] [Google Scholar]
  17. Kuehn M. J., Heuser J., Normark S., Hultgren S. J. P pili in uropathogenic E. coli are composite fibres with distinct fibrillar adhesive tips. Nature. 1992 Mar 19;356(6366):252–255. doi: 10.1038/356252a0. [DOI] [PubMed] [Google Scholar]
  18. Maki S., Kuribayashi M., Miki T., Horiuchi T. An amber replication mutant of F plasmid mapped in the minimal replication region. Mol Gen Genet. 1983;191(2):231–237. doi: 10.1007/BF00334819. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. McCormick B. A., Franklin D. P., Laux D. C., Cohen P. S. Type 1 pili are not necessary for colonization of the streptomycin-treated mouse large intestine by type 1-piliated Escherichia coli F-18 and E. coli K-12. Infect Immun. 1989 Oct;57(10):3022–3029. doi: 10.1128/iai.57.10.3022-3029.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Orndorff P. E., Falkow S. Nucleotide sequence of pilA, the gene encoding the structural component of type 1 pili in Escherichia coli. J Bacteriol. 1985 Apr;162(1):454–457. doi: 10.1128/jb.162.1.454-457.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Orndorff P. E., Falkow S. Organization and expression of genes responsible for type 1 piliation in Escherichia coli. J Bacteriol. 1984 Aug;159(2):736–744. doi: 10.1128/jb.159.2.736-744.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Ponniah S., Endres R. O., Hasty D. L., Abraham S. N. Fragmentation of Escherichia coli type 1 fimbriae exposes cryptic D-mannose-binding sites. J Bacteriol. 1991 Jul;173(13):4195–4202. doi: 10.1128/jb.173.13.4195-4202.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schwan W. R., Seifert H. S., Duncan J. L. Growth conditions mediate differential transcription of fim genes involved in phase variation of type 1 pili. J Bacteriol. 1992 Apr;174(7):2367–2375. doi: 10.1128/jb.174.7.2367-2375.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Spears P. A., Schauer D., Orndorff P. E. Metastable regulation of type 1 piliation in Escherichia coli and isolation and characterization of a phenotypically stable mutant. J Bacteriol. 1986 Oct;168(1):179–185. doi: 10.1128/jb.168.1.179-185.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES