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H-pilus assembly kinetics determined by electron microscopy

D Maher1, R Sherburne, D E Taylor

  • 1Department of Medical Microbiology and Infectious Diseases, University of Alberta, Edmonton, Canada.

Journal of Bacteriology
|April 1, 1993
PubMed
Summary
This summary is machine-generated.

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Escherichia coli pili extend by extrusion, not tip addition. Full-length pili regrow in 20 minutes, but high temperatures cause pili to vanish, inhibiting bacterial conjugation.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • IncHI1 plasmids, like R27, are crucial for bacterial conjugation and exhibit thermosensitive transfer properties.
  • Understanding pilus dynamics in Escherichia coli is key to deciphering bacterial conjugation mechanisms.

Purpose of the Study:

  • To investigate the kinetics of pilus outgrowth and retraction in Escherichia coli.
  • To elucidate the structural dynamics of H pili under varying temperature conditions.

Main Methods:

  • Utilized transmission electron microscopy (TEM) with bacteriophage labeling to visualize H pili.
  • Employed field emission scanning electron microscopy (FESEM) to observe pilus morphology.
  • Assessed pilus length and regrowth rates after mechanical removal (vortexing).

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Main Results:

  • H pili extend via cell surface extrusion, not pilin addition at the tip.
  • Full-length pili (2 microns) required 20 minutes for outgrowth after vortexing.
  • Pili retracted rapidly (within 1 minute) at 50°C, inhibiting mating aggregate formation at 37°C.

Conclusions:

  • Pilus extrusion is the primary mechanism for H pilus extension in Escherichia coli.
  • Temperature significantly impacts pilus stability and function, affecting bacterial conjugation.
  • The dynamic nature of pili suggests a role beyond simple adhesion, potentially in cell-cell interactions during conjugation.