Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Jan;169(1):157–163. doi: 10.1128/jb.169.1.157-163.1987

Immunoelectron microscopic analysis of elongation of type 1 fimbriae in Escherichia coli.

M A Lowe, S C Holt, B I Eisenstein
PMCID: PMC211747  PMID: 2878917

Abstract

Using 10- and 20-nm-diameter gold particles conjugated to an antifimbrial monoclonal antibody, we analyzed the location of assembly of newly formed subunits on growing type 1 fimbriae of Escherichia coli. Fimbriae were removed from an E. coli K-12-derived strain, CSH50, by blending. Blended cells were allowed to regenerate their fimbriae in growth medium for approximately 25 min, after which they were labeled with a 20-nm-gold-monoclonal antibody probe. Continued outgrowth of these labeled fimbriae was allowed for additional time intervals, after which they were labeled with a 10-nm-gold-monoclonal antibody probe. The resulting fimbriae, double labeled with 10- and 20-nm-diameter gold particles, were examined in an electron microscope. The pattern of labeling on individual fimbrial organelles indicated morphologically that newly synthesized subunits are added to a growing organelle at its base.

Full text

PDF
158

Images in this article

159Image
on p.159
160Image
on p.160
161Image
on p.161
162Image
on p.162

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. Brinton C. C., Jr The structure, function, synthesis and genetic control of bacterial pili and a molecular model for DNA and RNA transport in gram negative bacteria. Trans N Y Acad Sci. 1965 Jun;27(8):1003–1054. doi: 10.1111/j.2164-0947.1965.tb02342.x. [DOI] [PubMed] [Google Scholar]
  3. Dodd D. C., Bassford P. J., Jr, Eisenstein B. I. Dependence of secretion and assembly of type 1 fimbrial subunits of Escherichia coli on normal protein export. J Bacteriol. 1984 Sep;159(3):1077–1079. doi: 10.1128/jb.159.3.1077-1079.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dodd D. C., Eisenstein B. I. Antigenic quantitation of type 1 fimbriae on the surface of Escherichia coli cells by an enzyme-linked immunosorbent inhibition assay. Infect Immun. 1982 Nov;38(2):764–773. doi: 10.1128/iai.38.2.764-773.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dodd D. C., Eisenstein B. I. Kinetic analysis of the synthesis and assembly of type 1 fimbriae of Escherichia coli. J Bacteriol. 1984 Oct;160(1):227–232. doi: 10.1128/jb.160.1.227-232.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Emerson S. U., Tokuyasu K., Simon M. I. Bacterial flagella: polarity of elongation. Science. 1970 Jul 10;169(3941):190–192. doi: 10.1126/science.169.3941.190. [DOI] [PubMed] [Google Scholar]
  8. Eshdat Y., Silverblatt F. J., Sharon N. Dissociation and reassembly of Escherichia coli type 1 pili. J Bacteriol. 1981 Oct;148(1):308–314. doi: 10.1128/jb.148.1.308-314.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ey P. L., Prowse S. J., Jenkin C. R. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-sepharose. Immunochemistry. 1978 Jul;15(7):429–436. doi: 10.1016/0161-5890(78)90070-6. [DOI] [PubMed] [Google Scholar]
  10. Iino T. Polarity of flagellar growth in salmonella. J Gen Microbiol. 1969 May;56(2):227–239. doi: 10.1099/00221287-56-2-227. [DOI] [PubMed] [Google Scholar]
  11. Klemm P. The fimA gene encoding the type-1 fimbrial subunit of Escherichia coli. Nucleotide sequence and primary structure of the protein. Eur J Biochem. 1984 Sep 3;143(2):395–399. doi: 10.1111/j.1432-1033.1984.tb08386.x. [DOI] [PubMed] [Google Scholar]
  12. McMichael J. C., Ou J. T. Structure of common pili from Escherichia coli. J Bacteriol. 1979 Jun;138(3):969–975. doi: 10.1128/jb.138.3.969-975.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Novotny C., Carnahan J., Brinton C. C., Jr Mechanical removal of F pili, type I pili, and flagella from Hfr and RTF donor cells and the kinetics of their reappearance. J Bacteriol. 1969 Jun;98(3):1294–1306. doi: 10.1128/jb.98.3.1294-1306.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Oliver D. Protein secretion in Escherichia coli. Annu Rev Microbiol. 1985;39:615–648. doi: 10.1146/annurev.mi.39.100185.003151. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Randall L. L., Hardy S. J. Export of protein in bacteria. Microbiol Rev. 1984 Dec;48(4):290–298. doi: 10.1128/mr.48.4.290-298.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Robinson E. N., Jr, McGee Z. A., Kaplan J., Hammond M. E., Larson J. K., Buchanan T. M., Schoolnik G. K. Ultrastructural localization of specific gonococcal macromolecules with antibody-gold sphere immunological probes. Infect Immun. 1984 Nov;46(2):361–366. doi: 10.1128/iai.46.2.361-366.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Salit I. E., Gotschlich E. C. Type I Escherichia coli pili: characterization of binding to monkey kidney cells. J Exp Med. 1977 Nov 1;146(5):1182–1194. doi: 10.1084/jem.146.5.1182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sleytr U. B., Glauert A. M. Evidence for an empty core in a bacterial flagellum. Nature. 1973 Feb 23;241(5391):542–543. doi: 10.1038/241542a0. [DOI] [PubMed] [Google Scholar]
  20. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Zieg J., Simon M. Analysis of the nucleotide sequence of an invertible controlling element. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4196–4200. doi: 10.1073/pnas.77.7.4196. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES