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. 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.

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Selected References

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  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]

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