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Related Experiment Videos

A force-dependent switch reverses type IV pilus retraction.

Berenike Maier1, Michael Koomey, Michael P Sheetz

  • 1Department of Biological Sciences, Columbia University, 1212 Amsterdam Avenue, New York, NY 10027.

Proceedings of the National Academy of Sciences of the United States of America
|July 17, 2004
PubMed
Summary
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Type IV pilus retraction in Neisseria gonorrhoeae is usually irreversible. However, reduced PilT levels allow external forces to induce significant pilus elongation, reversing the retraction mechanism.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biophysics

Background:

  • Type IV pili are crucial for prokaryotic virulence, motility, and DNA transfer.
  • The type IV pilus system functions as a molecular machine for transporting polymers and DNA across the bacterial cell envelope.
  • In Neisseria gonorrhoeae, pilus retraction is primarily mediated by PilT, an AAA ATPase, and is considered irreversible.

Purpose of the Study:

  • To investigate the dynamics of type IV pilus under varying external forces.
  • To explore the mechanism of pilus elongation when retraction is impaired.
  • To model the force-dependent regulation of pilus dynamics.

Main Methods:

  • Utilizing atomic force microscopy to apply controlled forces to single pili.
  • Manipulating PilT protein levels in Neisseria gonorrhoeae.

Related Experiment Videos

  • Employing chemical kinetics modeling to analyze pilus dynamics.

    • Main Results:

    • Reduced PilT levels enable processive pilus elongation under high external forces (110 +/- 10 pN).
    • Pilus elongation occurs at a rate of 350 +/- 50 nm/s at forces >50 pN.
    • Elongation velocity is force-dependent below 50 pN, with relaxation causing immediate retraction.

    Conclusions:

    • A force-dependent molecular switch can reverse the pilus retraction mechanism, inducing elongation.
    • Pilus retraction and force-induced elongation share the same step length in the rate-limiting translocation step.
    • These findings provide insights into the mechanical regulation of type IV pilus systems.