Integrins in focal adhesions (FAs) mediate adhesion and force transmission to extracellular matrices essential for cell motility, proliferation and differentiation. Different fibronectin-binding integrins, simultaneously present in FAs, perform distinct functions. Yet, how integrin dynamics control biochemical and biomechanical processes in FAs is still elusive. Using single-protein tracking and super-resolution imaging we revealed the dynamic nano-organizations of integrins and talin inside FAs. Integrins reside in FAs through free-diffusion and immobilization cycles. Integrin activation promotes immobilization, stabilized in FAs by simultaneous connection to fibronectin and actin-binding proteins. Talin is recruited in FAs directly from the cytosol without membrane free-diffusion, restricting integrin immobilization to FAs. Immobilized β3-integrins are enriched and stationary within FAs, whereas immobilized β1-integrins are less enriched and exhibit rearward movements. Talin is enriched and mainly stationary, but also exhibited rearward movements in FAs, consistent with stable connections with both β-integrins. Thus, differential transmission of actin motion to fibronectin occurs through specific integrins within FAs.
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We thank C. Breillat, A. Frouin, D. Bouchet and P. Gonzales for technical assistance; M. P. Sheetz, O. Thoumine, J. Petersen, B. Fourcade, M. Block, L. Duchesne and D.G. Fernig for helpful discussions; M. Humphries, N. Kieffer, J. Wehland, A. Gautreau and P. Kanchanawong for the gift of reagents; P. Legros and C. Poujol (Bordeaux Imaging Center) for STED imaging. We acknowledge financial support from the French Ministry of Research and CNRS, ANR grant Nanomotility (G.G., B.L., O.R.), Fondation ARC pour la Recherche sur le Cancer (O.R.), Conseil Régional Aquitaine, Fondation pour la Recherche Médicale, the ERC Program numbers 232942 Nano-Dyn-Syn (D.C., B.L.) and 235552 Glutraf (D.N.), the Human Frontiers Science Programme (B.L.) and The Ligue National contre le Cancer—équipe labellisée 2010 (C.A-R., O.D.). The research was conducted in the scope of the International Associated Laboratory LIA CAFS.
The authors declare no competing financial interests.
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Rossier, O., Octeau, V., Sibarita, JB. et al. Integrins β1 and β3 exhibit distinct dynamic nanoscale organizations inside focal adhesions. Nat Cell Biol 14, 1057–1067 (2012). https://doi.org/10.1038/ncb2588
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