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. 2001 Jun 5;98(12):6901-4.
doi: 10.1073/pnas.121171698. Epub 2001 May 29.

Direct observation of extension and retraction of type IV pili

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Direct observation of extension and retraction of type IV pili

J M Skerker et al. Proc Natl Acad Sci U S A. .

Abstract

Type IV pili are thin filaments that extend from the poles of a diverse group of bacteria, enabling them to move at speeds of a few tenths of a micrometer per second. They are required for twitching motility, e.g., in Pseudomonas aeruginosa and Neisseria gonorrhoeae, and for social gliding motility in Myxococcus xanthus. Here we report direct observation of extension and retraction of type IV pili in P. aeruginosa. Cells without flagellar filaments were labeled with an amino-specific Cy3 fluorescent dye and were visualized on a quartz slide by total internal reflection microscopy. When pili were attached to a cell and their distal ends were free, they extended or retracted at rates of about 0.5 microm s(-1) (29 degrees C). They also flexed by Brownian motion, exhibiting a persistence length of about 5 microm. Frequently, the distal tip of a filament adsorbed to the substratum and the filament was pulled taut. From the absence of lateral deflections of such filaments, we estimate tensions of at least 10 pN. Occasionally, cell bodies came free and were pulled forward by pilus retraction. Thus, type IV pili are linear actuators that extend, attach at their distal tips, exert substantial force, and retract.

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Figures

Figure 1
Figure 1
Visualization of Cy3-labeled pili on nonflagellate cells of P. aeruginosa. (A and B) Filament extension and retraction. (C) Filaments under tension. (D and E) Cell movement. Elapsed time (t) in s. (Bars = 2 μm.)
Figure 2
Figure 2
A broken pilus exhibiting thermal fluctuations in shape over a time span of 15 s. This filament was attached to the substratum at its left end. Elapsed time (t) in s. (Bar = 2 μm.)
Figure 3
Figure 3
Histograms of rates of pilus extension and retraction. (A) Extension of 17 pili. (B) Retraction of 57 pili.

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