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Forces and constraints controlling podosome assembly and disassembly

Nisha Mohd Rafiq, Gianluca Grenci, Michael M. Kozlov, Gareth E Jones, Virgile Viasnoff, Alexander D. Bershadsky
doi: https://doi.org/10.1101/495176
Nisha Mohd Rafiq
1Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore
2Randall Centre for Cell & Molecular Biophysics, King’s College London, London SE14 5TG, UK
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Gianluca Grenci
1Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore
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Michael M. Kozlov
3Department of Physiology and Pharmacology, Sackler Faculty of Medicine, Tel Aviv University, 69978 Tel Aviv, Israel
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Gareth E Jones
2Randall Centre for Cell & Molecular Biophysics, King’s College London, London SE14 5TG, UK
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Virgile Viasnoff
1Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore
4CNRS UMI 3639, 5A engineering drive 1, 117411 Singapore
5Department of Biological Sciences, National university of Singapore, 14 Science Drive 4, Singapore 117543
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Alexander D. Bershadsky
1Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore
6Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel
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Abstract

Podosomes are a singular category of integrin-mediated adhesions important in the processes of cell migration, matrix degradation, and cancer cell invasion. Despite a wealth of biochemical studies, the effects of mechanical forces on podosome integrity and dynamics are poorly understood. Here, we show that podosomes are highly sensitive to two groups of physical factors. First, we describe the process of podosome disassembly induced by activation of myosin-IIA filament assembly. Next, we find that podosome integrity and dynamics depends upon membrane tension and can be experimentally perturbed by osmotic swelling and deoxycholate treatment. We have also found that podosomes can be disrupted in a reversible manner by single or cyclic radial stretching of the substratum. We show that disruption of podosomes induced by osmotic swelling is independent of myosin-II filaments. Inhibition of the membrane sculpting protein, dynamin-II, but not clathrin, resulted in activation of myosin-IIA filament formation and disruption of podosomes. The effect of dynamin-II inhibition on podosomes was however independent of myosin-II filaments. Moreover, formation of organized arrays of podosomes in response to microtopographic cues (the ridges with triangular profile) was not accompanied by reorganization of myosin-II filaments. Thus, mechanical elements such as myosin-II filaments and factors affecting membrane tension/sculpting independently modulate podosome formation and dynamics, underlying a versatile response of these adhesion structures to intracellular and extracellular cues.

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Posted December 13, 2018.
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Forces and constraints controlling podosome assembly and disassembly
Nisha Mohd Rafiq, Gianluca Grenci, Michael M. Kozlov, Gareth E Jones, Virgile Viasnoff, Alexander D. Bershadsky
bioRxiv 495176; doi: https://doi.org/10.1101/495176
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Forces and constraints controlling podosome assembly and disassembly
Nisha Mohd Rafiq, Gianluca Grenci, Michael M. Kozlov, Gareth E Jones, Virgile Viasnoff, Alexander D. Bershadsky
bioRxiv 495176; doi: https://doi.org/10.1101/495176

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