Abstract
Prokaryotes have the ability to walk on surfaces using type IV pili (TFP), a motility mechanism known as twitching1,2. Molecular motors drive TFP extension and retraction, but whether and how these movements are coordinated is unknown3. Here, we reveal how the pathogen Pseudomonas aeruginosa coordinates the motorized activity of TFP to power efficient surface motility. To do this, we dynamically visualized TFP extension, attachment and retraction events at high resolution in four dimensions using label-free interferometric scattering microscopy (iSCAT)4. By measuring TFP dynamics, we found that the retraction motor PilT was sufficient to generate tension and power motility in free solution, while its partner ATPase PilU may improve retraction only in high-friction environments. Using precise timing of successive attachment and retraction, we show that P. aeruginosa engages PilT motors very rapidly and almost only when TFP encounter the surface, suggesting contact sensing. Finally, measurements of TFP dwell times on surfaces show that tension reinforced the adhesion strength to the surface of individual pili, thereby increasing effective pulling time during retraction. The successive control of TFP extension, attachment, retraction and detachment suggests that sequential control of motility machinery is a conserved strategy for optimized locomotion across domains of life.
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Code availability
All codes are available from the corresponding author upon reasonable request.
Data availability
All data are available from the corresponding author upon reasonable request.
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Acknowledgements
The authors would like to thank J. Andrecka for valuable discussions on iSCAT, J. Engel and Y. Inclan for strains and plasmids and Z. Al-Mayyah for help with generating one mutant strain. L.T. and A.P. thank the Swiss National Foundation for funding this work through Projects (grant No. 31003A_169377) and the Gabriella Giorgi-Cavaglieri Foundation.
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L.T. and A.P conceptualized the study and performed experiments and data analysis. L.T., A.F. and P.K. implemented and adapted the iSCAT microscope for live-cell imaging. L.T., P.K and A.P. wrote the manuscript.
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Supplementary information
Supplementary Information
Supplementary Figures 1–7 and Supplementary Video Legends.
Supplementary Video 1
iSCAT visualization of a twitching WT cell.
Supplementary Video 2
iSCAT visualization of a twitching fliC cell.
Supplementary Video 3
iSCAT visualization of the fliC mutant.
Supplementary Video 4
iSCAT visualization of the pilTU fliC mutant.
Supplementary Video 5
iSCAT visualization of the retraction motor mutant pilT fliC.
Supplementary Video 6
iSCAT visualization of the retraction motor mutant pilU fliC.
Supplementary Video 7
iSCAT visualization of a twitching pilU fliC cell.
Supplementary Video 8
Visualization of twitching motility in high friction environment highlights a function for PilU in force generation.
Supplementary Video 9
iSCAT visualization of extension, attachment and retraction of a TFP.
Supplementary Video 10
iSCAT visualization of extension, attachment and retraction of a TFP.
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Talà, L., Fineberg, A., Kukura, P. et al. Pseudomonas aeruginosa orchestrates twitching motility by sequential control of type IV pili movements. Nat Microbiol 4, 774–780 (2019). https://doi.org/10.1038/s41564-019-0378-9
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DOI: https://doi.org/10.1038/s41564-019-0378-9
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