Abstract
Mechanical forces on matrix–integrin–cytoskeleton linkages are crucial for cell viability, morphology and organ function1. The production of force depends on the molecular connections from extracellular-matrix–integrin complexes to the cytoskeleton2,3. The minimal matrix complex causing integrin–cytoskeleton connections is a trimer of fibronectin's integrin-binding domain FNIII7-10 (ref. 4). Here we report a specific, molecular slip bond that was broken repeatedly by a force of 2 pN at the cellular loading rate of 60 nm s-1; this occurred with single trimer beads but not with monomer. Talin1, which binds to both integrins and actin filaments in vitro, is required for the 2-pN slip bond and rapid cytoskeleton binding. Further, inhibition of fibronectin binding to αvβ3 and deletion of β3 markedly decreases the 2-pN force peak. We suggest that talin1 initially forms a molecular slip bond between closely packed fibronectin–integrin complexes and the actin cytoskeleton, which can apply a low level of force to fibronectin until many bonds form or a signal is received to activate a force response.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Geiger, B. & Bershadsky, A. Assembly and mechanosensory function of focal contacts. Curr. Opin. Cell Biol. 13, 584–592 (2001)
Critchley, D. R. Focal adhesions—the cytoskeletal connection. Curr. Opin. Cell Biol. 12, 133–139 (2000)
Sheetz, M. P., Felsenfeld, D. P. & Galbraith, C. G. Cell migration: Regulation of force on extracellular–matrix–integrin complexes. Trends Cell Biol. 8, 51–54 (1998)
Coussen, F., Choquet, D., Sheetz, M. P. & Erickson, H. P. Trimers of the fibronectin cell adhesion domain localize to actin filament bundles and undergo rearward translocation. J. Cell Sci. 115, 2581–2590 (2002)
Lauffenburger, D. A. & Horwitz, A. F. Cell migration: A physically integrated molecular process. Cell 84, 359–369 (1996)
Riveline, D. et al. Focal contacts as mechanosensors: Externally applied local mechanical force induces growth of focal contacts by an mDia1-dependent and ROCK- independent mechanism. J. Cell Biol. 153, 1175–1186 (2001)
Galbraith, C. G., Yamada, K. M. & Sheetz, M. P. The relationship between force and focal complex development. J. Cell Biol. 159, 695–705 (2002)
Choquet, D., Felsenfeld, D. P. & Sheetz, M. P. Extracellular matrix rigidity causes strengthening of integrin–cytoskeleton linkages. Cell 88, 39–48 (1997)
Sonnenberg, A. Integrins and their ligands. Curr. Top. Microbiol. Immunol. 184, 7–35 (1993)
Pierschbacher, M. D. & Ruoslahti, E. Influence of stereochemistry of the sequence Arg-Gly-Asp-Xaa on binding specificity in cell adhesion. J. Biol. Chem. 262, 17294–17298 (1987)
Thoumine, O., Kocian, P., Kottelat, A. & Meister, J. J. Short-term binding of fibroblasts to fibronectin: Optical tweezers experiments and probabilistic analysis. Eur. Biophys. J. 29, 398–408 (2000)
Litvinov, R. I., Shuman, H., Bennett, J. S. & Weisel, J. W. Binding strength and activation state of single fibrinogen–integrin pairs on living cells. Proc. Natl Acad. Sci. USA 99, 7426–7431 (2002)
Finer, J. T., Mehta, A. D. & Spudich, J. A. Characterization of single actin–myosin interactions. Biophys. J. 68, 291S–296S (1995)
Nishizaka, T., Miyata, H., Yoshikawa, H., Ishiwata, S. & Kinosita, K. Jr Unbinding force of a single motor molecule of muscle measured using optical tweezers. Nature 377, 251–254 (1995)
Suzuki, K. & Sheetz, M. P. Binding of cross-linked glycosylphosphatidylinositol-anchored proteins to discrete actin-associated sites and cholesterol-dependent domains. Biophys. J. 81, 2181–2189 (2001)
Giannone, G., Jiang, G., Sutton, D. H., Critchley, D. R. & Sheetz, M. P. Talin1 is essential for force-dependent reinforcement of integrin–cytoskeleton connections but not for activation of tyrosine kinases. J. Cell Biol. (submitted)
Calderwood, D. A. et al. The Talin head domain binds to integrin beta subunit cytoplasmic tails and regulates integrin activation. J. Biol. Chem. 274, 28071–28074 (1999)
Brown, N. H. et al. Talin is essential for integrin function in Drosophila. Dev. Cell 3, 569–579 (2002)
Nishizaka, T., Shi, Q. & Sheetz, M. P. Position-dependent linkages of fibronectin–integrin–cytoskeleton. Proc. Natl Acad. Sci. USA 97, 692–697 (2000)
von Wichert, G. et al. RPTP-α acts as a transducer of mechanical force on αv/β3–integrin–cytoskeleton linkages. J. Cell Biol. 161, 143–153 (2003)
Su, J., Muranjan, M. & Sap, J. Receptor protein tyrosine phosphatase α activates Src-family kinases and controls integrin-mediated responses in fibroblasts. Curr. Biol. 9, 505–511 (1999)
Balaban, N. Q. et al. Force and focal adhesion assembly: A close relationship studied using elastic micropatterned substrates. Nature Cell Biol. 3, 466–472 (2001)
Watanabe, K. et al. Molecular mechanics of cardiac titin's PEVK and N2B spring elements. J. Biol. Chem. 277, 11549–11558 (2002)
Oberhauser, A. F., Marszalek, P. E., Erickson, H. P. & Fernandez, J. M. The molecular elasticity of the extracellular matrix protein tenascin. Nature 393, 181–185 (1998)
Carrion-Vazquez, M. et al. Mechanical and chemical unfolding of a single protein: A comparison. Proc. Natl Acad. Sci. USA 96, 3694–3699 (1999)
Merkel, R., Nassoy, P., Leung, A., Ritchie, K. & Evans, E. Energy landscapes of receptor–ligand bonds explored with dynamic force spectroscopy. Nature 397, 50–53 (1999)
Xing, B., Jedsadayanmata, A. & Lam, S. C. Localization of an integrin binding site to the C terminus of talin. J. Biol. Chem. 276, 44373–44378 (2001)
Hodivala-Dilke, K. M. et al. β3-integrin-deficient mice are a model for Glanzmann thrombasthenia showing placental defects and reduced survival. J. Clin. Invest. 103, 229–238 (1999)
Priddle, H. et al. Disruption of the talin gene compromises focal adhesion assembly in undifferentiated but not differentiated embryonic stem cells. J. Cell Biol. 142, 1121–1133 (1998)
Felsenfeld, D. P., Choquet, D. & Sheetz, M. P. Ligand binding regulates the directed movement of β1 integrins on fibroblasts. Nature 383, 438–440 (1996)
Acknowledgements
We thank H. Erickson for FNIII7-10 monomer and trimer constructs; A. J. Woods for the talin1-ABS construct; R. Hynes for β3 cell lines; J. Sap for RPTPα cell lines; G. von Wichert and N. Heckenberg for discussions; and all the Sheetz laboratory members for consistent help. This work was supported by a grant from the NIH to M.P.S. Work in D.R.C.'s laboratory was funded by the Wellcome Trust.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare that they have no competing financial interests.
Rights and permissions
About this article
Cite this article
Jiang, G., Giannone, G., Critchley, D. et al. Two-piconewton slip bond between fibronectin and the cytoskeleton depends on talin. Nature 424, 334–337 (2003). https://doi.org/10.1038/nature01805
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nature01805
This article is cited by
-
Quantifying force transmission through fibroblasts: changes of traction forces under external shearing
European Biophysics Journal (2022)
-
The force loading rate drives cell mechanosensing through both reinforcement and cytoskeletal softening
Nature Communications (2021)
-
Forces generated by lamellipodial actin filament elongation regulate the WAVE complex during cell migration
Nature Cell Biology (2021)
-
Molecular motion and tridimensional nanoscale localization of kindlin control integrin activation in focal adhesions
Nature Communications (2021)
-
Kindlin3 regulates biophysical properties and mechanics of membrane to cortex attachment
Cellular and Molecular Life Sciences (2021)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.