Integrins β1 and β3 exhibit distinct dynamic nanoscale organizations inside focal adhesions

A Corrigendum to this article was published on 06 November 2012


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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Figure 1: β3-integrins are immobilized more frequently and undergo slower free-diffusion inside versus outside FAs.
Figure 2: Integrin immobilization correlates with integrin activation and require both fibronectin and ABPs binding.
Figure 3: Integrins undergo repeated cycles of slow free-diffusion and immobilization within FAs.
Figure 4: Talin is recruited in FAs directly from the cytosol without membrane free-diffusion, spatially restricting integrin immobilization to FAs.
Figure 5: Differential nanoscale organization of β3- and β1-integrins in FAs.
Figure 6: β3- and β1-integrin extracellular domains determine their distinct dynamic nanoscale organizations inside FAs.
Figure 7: Differential transmission of F-actin motion to β3-integrin, β1-integrin and talin.


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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.

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O.R. and G.G. conceptualized and performed the sptPALM and STED experiments. V.O., C.L., L.C. and B.L. conceptualized and developed the single-protein-tracking set-up used for ATTO647N. G.G. performed single-protein-tracking experiments using ATTO647N. B.T. designed and generated new protein constructs. V.G. and R.T. designed and synthesized the TrisNTA–ATTO647N. D.N. and J-B.S. developed the sptPALM set-up. J-B.S. developed the analytical tools for sptPALM. V.O., C.L. and L.C. developed the analytical tools for single-protein tracking using ATTO647N. O.D. and C.A-R. contributed valuable scientific advice and developed the β1-integrin–mEOS2 and chimaeric integrin constructs. O.D. performed FACS experiments. L.C., D.C., B.L. and G.G. coordinated the study. O.R. and G.G. wrote the manuscript and Supplementary Information. All authors discussed the results and commented on the manuscript.

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Correspondence to Grégory Giannone.

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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).

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