Abstract
Twitching and social gliding motility allow many Gram negative bacteria to crawl along surfaces, and are implicated in a wide range of biological functions1. Type IV pili (Tfp) are required for twitching and social gliding, but the mechanism by which these filaments promote motility has remained enigmatic1,2,3,4. Here we use laser tweezers5 to show that Tfp forcefully retract. Neisseria gonorrhoeae cells that produce Tfp actively crawl on a glass surface and form adherent microcolonies. When laser tweezers are used to place and hold cells near a microcolony, retractile forces pull the cells toward the microcolony. In quantitative experiments, the Tfp of immobilized bacteria bind to latex beads and retract, pulling beads from the tweezers at forces that can exceed 80 pN. Episodes of retraction terminate with release or breakage of the Tfp tether. Both motility and retraction mediated by Tfp occur at about 1 µm s-1 and require protein synthesis and function of the PilT protein. Our experiments establish that Tfp filaments retract, generate substantial force and directly mediate cell movement.
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References
Wall, D. & Kaiser, D. Type IV pili and cell motility. Mol. Microbiol. 32, 1–10 (1999).
Henrichsen, J. Twitching motility. Annu. Rev. Microbiol. 37, 81–93 (1983).
Bradley, D. E. A function of Pseudomonas aeruginosa PAO polar pili: twitching motility. Can. J. Microbiol. 26, 146– 154 (1980).
Wolfgang, M., Park, H. S., Hayes, S. F., van Putten, J. P. M. & Koomey, M. Suppression of an absolute defect in type IV pilus biogenesis by loss-of-function mutations in pilT, a twitching motility gene in Neisseria gonorrhoeae. Proc. Natl Acad. Sci. USA 95, 14973– 14978 (1998).
Sheetz, M. P. (ed.) Laser Tweezers in Cell Biology (Academic, New York, 1997).
Swanson, J. Studies on gonococcus infection. XII. Colony color and opacity variants of gonococci. Infect. Immun. 19, 320– 331 (1978).
O'Toole, G. A. & Kolter, R. Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol. Microbiol. 30, 295–304 (1998).
Bieber, D. et al. Type IV pili, transient bacterial aggregates, and virulence of enteropathogenic Escherichia coli. Science 280 , 2114–2118 (1998).
Comolli, J. C. et al. Pseudomonas aeruginosa gene products PilT and PilU are required for cytotoxicity in vitro and virulence in a mouse model of acute pneumonia. Infect. Immun. 67, 3625– 3630 (1999).
Pujol, C., Eugene, E., Marceau, M. & Nassif, X. The meningococcal PilT protein is required for induction of intimate attachment to epithelial cells following pilus-mediated adhesion. Proc. Natl Acad. Sci. USA 96, 4017–4022 ( 1999).
Merz, A. J., Enns, C. A. & So, M. Type IV pili of pathogenic Neisseriae elicit cortical plaque formation in epithelial cells. Mol. Microbiol. 32, 1316–1332 (1999).
Seifert, H. S., Ajioka, R. S., Marchal, C., Sparling, P. F. & So, M. DNA transformation leads to pilin antigenic variation in Neisseria gonorrhoeae. Nature 336 , 392–395 (1988).
Dubnau, D. DNA uptake in bacteria. Annu. Rev. Microbiol. 53, 217–244 (1999).
Yoshida, T., Kim, S. R. & Komano, T. Twelve pil genes are required for biogenesis of the R64 thin pilus. J. Bacteriol. 181, 2038–2043 (1999).
Bradley, D. E. Evidence for the retraction of Pseudomonas aeruginosa RNA phage pili. Biochem. Biophys. Res. Commun. 47, 142– 149 (1972).
Karaolis, D. K., Somara, S., Maneval, D. R. Jr, Johnson, J. A. & Kaper, J. B. A bacteriophage encoding a pathogenicity island, a type-IV pilus and a phage receptor in cholera bacteria. Nature 399, 375– 379 (1999).
Parge, H. E. et al. Structure of the fibre-forming protein pilin at 2.6 Å resolution. Nature 378, 32– 38 (1995).
Forest, K. T. & Tainer, J. A. Type-4 pilus structure: outside to inside and top to bottom—a minireview. Gene 192, 165–169 (1997).
Whitchurch, C. B., Hobbs, M., Livingston, S. P., Krishnapillai, V. & Mattick, J. S. Characterisation of a Pseudomonas aeruginosa twitching motility gene and evidence for a specialised protein export system widespread in eubacteria. Gene 101, 33–44 (1991).
Wolfgang, M. et al. pilT mutations lead to simultaneous defects in competence for natural transformation and twitching motility in piliated Neisseria gonorrhoeae. Mol. Microbiol. 29, 321 –330 (1998).
Brossay, L., Paradis, G., Fox, R., Koomey, M. & Hebert, J. Identification, localization, and distribution of the PilT protein in Neisseria gonorrhoeae. Infect. Immun. 62, 2302–2308 (1994).
Krause, S. et al. Sequence-related protein export NTPases encoded by the conjugative transfer region of RP4 and by the cag pathogenicity island of Helicobacter pylori share similar hexameric ring structures. Proc. Natl Acad. Sci. USA 97, 3067–3072 (2000).
Novotny, C. P. & Fives-Taylor, P. Retraction of F pili. J. Bacteriol. 117, 1306– 1311 (1974).
Ginocchio, C. C., Olmsted, S. B., Wells, C. L. & Galan, J. E. Contact with epithelial cells induces the formation of surface appendages on Salmonella typhimurium. Cell 76, 717–724 (1994).
Evans, E., Berk, D. & Leung, A. Detachment of agglutinin-bonded red blood cells. I. Forces to rupture molecular-point attachments. Biophys. J. 59, 838– 848 (1991).
Shao, J. Y., Ting-Beall, H. P. & Hochmuth, R. M. Static and dynamic lengths of neutrophil microvilli. Proc. Natl Acad. Sci. USA 95, 6797– 6802 (1998).
Coppin, C. M., Finer, J. T., Spudich, J. A. & Vale, R. D. Detection of sub-8-nm movements of kinesin by high-resolution optical-trap microscopy. Proc. Natl Acad. Sci. USA 93, 1913–1917 (1996).
Mahadevan, L. & Matsudaira, P. Motility powered by supramolecular springs and ratchets. Science 288, 95– 100 (2000).
Dupuy, B., Taha, M. K., Pugsley, A. P. & Marchal, C. Neisseria gonorrhoeae prepilin export studied in Escherichia coli. J. Bacteriol. 173, 7589– 7598 (1991).
Felsenfeld, D. P., Schwartzberg, P. L., Venegas, A., Tse, R. & Sheetz, M. P. Selective regulation of integrin-cytoskeleton interactions by the tyrosine kinase Src. Nature Cell Biol. 1, 200–206 (1999).
Acknowledgements
We thank our colleagues in the Sheetz and So labs for invaluable technical assistance and stimulating discussions; E. Barklis and L. Kenney for critical comments on the manuscript; and M. Koomey for providing bacterial strains. This work was supported by NIH grants to M.S. and M.P.S. A.J.M. received pre-doctoral support from an NIH NRSA grant and postdoctoral support from the Cancer Research Fund of the Damon Runyan-Walter Winchell Foundation.
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Merz, A., So, M. & Sheetz, M. Pilus retraction powers bacterial twitching motility. Nature 407, 98–102 (2000). https://doi.org/10.1038/35024105
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DOI: https://doi.org/10.1038/35024105
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