Extensive research over the past decades has identified integrins to be the primary transmembrane receptors that enable cells to respond to external mechanical cues. We reveal here a mechanism whereby syndecan-4 tunes cell mechanics in response to localized tension via a coordinated mechanochemical signalling response that involves activation of two other receptors: epidermal growth factor receptor and β1 integrin. Tension on syndecan-4 induces cell-wide activation of the kindlin-2/β1 integrin/RhoA axis in a PI3K-dependent manner. Furthermore, syndecan-4-mediated tension at the cell–extracellular matrix interface is required for yes-associated protein activation. Extracellular tension on syndecan-4 triggers a conformational change in the cytoplasmic domain, the variable region of which is indispensable for the mechanical adaptation to force, facilitating the assembly of a syndecan-4/α-actinin/F-actin molecular scaffold at the bead adhesion. This mechanotransduction pathway for syndecan-4 should have immediate implications for the broader field of mechanobiology.
Subscribe to Journal
Get full journal access for 1 year
only $16.58 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
The data that support the findings of this study are available from the corresponding authors upon reasonable request.
The MATLAB code used to track bead displacements in the magnetic tweezers experiments is available from A.E.d.R.H. upon reasonable request. Code used in the MD simulations is available from V.P.H. upon reasonable request.
Kim, C. W., Goldberger, O. A., Gallo, R. L. & Bernfield, M. Members of the syndecan family of heparan sulfate proteoglycans are expressed in distinct cell-, tissue- and development-specific patterns. Mol. Biol. Cell 5, 797–805 (1994).
Elfenbein, A. & Simons, M. Syndecan-4 signaling at a glance. J. Cell Sci. 126, 3799–3804 (2013).
Okina, E., Manon-Jensen, T., Whiteford, J. R. & Couchman, J. R. Syndecan proteoglycan contributions to cytoskeletal organization and contractility. Scand. J. Med. Sci. Sports 19, 479–489 (2009).
Morgan, M. R., Humphries, M. J. & Bass, M. D. Synergistic control of cell adhesion by integrins and syndecans. Nat. Rev. Mol. Cell Biol. 8, 957–969 (2007).
Saoncella, S. et al. Syndecan-4 signals cooperatively with integrins in a Rho-dependent manner in the assembly of focal adhesions and actin stress fibers. Proc. Natl Acad. Sci. USA 96, 2805–2810 (1999).
Fiore, V. F., Ju, L., Chen, Y., Zhu, C. & Barker, T. H. Dynamic catch of a Thy-1–α5β1+syndecan-4 trimolecular complex. Nat. Commun. 5, 4886 (2014).
Echtermeyer, F. et al. Delayed wound repair and impaired angiogenesis in mice lacking syndecan-4. J. Clin. Invest. 107, R9–R14 (2001).
Longley, R. L. et al. Control of morphology, cytoskeleton and migration by syndecan-4. J. Cell Sci. 112, 3421–3431 (1999).
Cavalheiro, R. P. et al. Coupling of vinculin to F-actin demands syndecan-4 proteoglycan. Matrix Biol. 63, 23–37 (2017).
Okina, E., Grossi, A., Gopal, S., Multhaupt, H. A. & Couchman, J. R. Alpha-actinin interactions with syndecan-4 are integral to fibroblast-matrix adhesion and regulate cytoskeletal architecture. Int. J. Biochem. Cell Biol. 44, 2161–2174 (2012).
Gopal, S. et al. Heparan sulfate chain valency controls syndecan-4 function in cell adhesion. J. Biol. Chem. 285, 14247–14258 (2010).
Chen, Y. et al. Matrix contraction by dermal fibroblasts requires transforming growth factor-β/activin-linked kinase 5, heparan sulfate-containing proteoglycans and MEK/ERK. Am. J. Pathol. 167, 1699–1711 (2005).
Florian, J. A. et al. Heparan sulfate proteoglycan is a mechanosensor on endothelial cells. Circ. Res. 93, e136–e142 (2003).
Moon, J. J. et al. Role of cell surface heparan sulfate proteoglycans in endothelial cell migration and mechanotransduction. J. Cell. Physiol. 203, 166–176 (2005).
Baeyens, N. et al. Syndecan 4 is required for endothelial alignment in flow and atheroprotective signaling. Proc. Natl Acad. Sci. USA 111, 17308–17313 (2014).
Wang, Y. et al. Syndecan 4 controls lymphatic vasculature remodeling during mouse embryonic development. Development 143, 4441–4451 (2016).
Bellin, R. M. et al. Defining the role of syndecan-4 in mechanotransduction using surface-modification approaches. Proc. Natl Acad. Sci. USA 106, 22102–22107 (2009).
Huang, C.-P., Cheng, C.-M., Su, H.-L. & Lin, Y.-W. Syndecan-4 promotes epithelial tumor cells spreading and regulates the turnover of PKCα activity under mechanical stimulation on the elastomeric substrates. Cell. Physiol. Biochem. 36, 1291–1304 (2015).
Guilluy, C. et al. The Rho GEFs LARG and GEF-H1 regulate the mechanical response to force on integrins. Nat. Cell Biol. 13, 722–727 (2011).
Collins, C. et al. Localized tensional forces on PECAM-1 elicit a global mechanotransduction response via the integrin-RhoA pathway. Curr. Biol. 22, 2087–2094 (2012).
Muhamed, I. et al. E-cadherin-mediated force transduction signals regulate global cell mechanics. J. Cell Sci. 129, 1843–1854 (2016).
Guilluy, C. et al. Isolated nuclei adapt to force and reveal a mechanotransduction pathway in the nucleus. Nat. Cell Biol. 16, 376–381 (2014).
Pierschbacher, M. D., Hayman, E. G. & Ruoslahti, E. Location of the cell-attachment site in fibronectin with monoclonal antibodies and proteolytic fragments of the molecule. Cell 26, 259–267 (1981).
Vanhaesebroeck, B., Guillermet-Guibert, J., Graupera, M. & Bilanges, B. The emerging mechanisms of isoform-specific PI3K signalling. Nat. Rev. Mol. Cell Biol. 11, 329–341 (2010).
Wang, H., Jin, H. & Rapraeger, A. C. Syndecan-1 and syndecan-4 capture epidermal growth factor receptor family members and the α3β1 integrin via binding sites in their ectodomains: novel synstatins prevent kinase capture and inhibit α6β4-integrin-dependent epithelial cell motility. J. Biol. Chem. 290, 26103–26113 (2015).
Lemmon, M. A. & Schlessinger, J. Cell signaling by receptor tyrosine kinases. Cell 141, 1117–1134 (2010).
Sakaguchi, M. et al. S100A11, an dual mediator for growth regulation of human keratinocytes. Mol. Biol. Cell 19, 78–85 (2008).
Muller-Deubert, S., Seefried, L., Krug, M., Jakob, F. & Ebert, R. Epidermal growth factor as a mechanosensitizer in human bone marrow stromal cells. Stem Cell Res. 24, 69–76 (2017).
Saxena, M. et al. EGFR and HER2 activate rigidity sensing only on rigid matrices. Nat. Mater. 16, 775–781 (2017).
Morgan, M. R. et al. Syndecan-4 phosphorylation is a control point for integrin recycling. Dev. Cell 24, 472–485 (2013).
Calderwood, D. A., Campbell, I. D. & Critchley, D. R. Talins and kindlins: partners in integrin-mediated adhesion. Nat. Rev. Mol. Cell Biol. 14, 503–517 (2013).
Liu, J. et al. Structural basis of phosphoinositide binding to kindlin-2 protein pleckstrin homology domain in regulating integrin activation. J. Biol. Chem. 286, 43334–43342 (2011).
Liu, Y., Zhu, Y., Ye, S. & Zhang, R. Crystal structure of kindlin-2 PH domain reveals a conformational transition for its membrane anchoring and regulation of integrin activation. Protein Cell 3, 434–440 (2012).
Qu, H. et al. Kindlin-2 regulates podocyte adhesion and fibronectin matrix deposition through interactions with phosphoinositides and integrins. J. Cell Sci. 124, 879–891 (2011).
Vining, K. H. & Mooney, D. J. Mechanical forces direct stem cell behaviour in development and regeneration. Nat. Rev. Mol. Cell Biol. 18, 728–742 (2017).
Panciera, T., Azzolin, L., Cordenonsi, M. & Piccolo, S. Mechanobiology of YAP and TAZ in physiology and disease. Nat. Rev. Mol. Cell Biol. 18, 758–770 (2017).
Hoffman, B. D., Grashoff, C. & Schwartz, M. A. Dynamic molecular processes mediate cellular mechanotransduction. Nature 475, 316–323 (2011).
Baietti, M. F. et al. Syndecan-syntenin-ALIX regulates the biogenesis of exosomes. Nat. Cell Biol. 14, 677–685 (2012).
Bass, M. D. & Humphries, M. J. Cytoplasmic interactions of syndecan-4 orchestrate adhesion receptor and growth factor receptor signalling. Biochem. J. 368, 1–15 (2002).
Dovas, A., Yoneda, A. & Couchman, J. R. PKCα-dependent activation of RhoA by syndecan-4 during focal adhesion formation. J. Cell Sci. 119, 2837–2846 (2006).
Greene, D. K., Tumova, S., Couchman, J. R. & Woods, A. Syndecan-4 associates with α-actinin. J. Biol. Chem. 278, 7617–7623 (2003).
Lim, S. T., Longley, R. L., Couchman, J. R. & Woods, A. Direct binding of syndecan-4 cytoplasmic domain to the catalytic domain of protein kinase C alpha (PKC alpha) increases focal adhesion localization of PKC alpha. J. Biol. Chem. 278, 13795–13802 (2003).
Roca-Cusachs, P. et al. Integrin-dependent force transmission to the extracellular matrix by α-actinin triggers adhesion maturation. Proc. Natl Acad. Sci. USA 110, E1361–E1370 (2013).
Tzima, E. et al. A mechanosensory complex that mediates the endothelial cell response to fluid shear stress. Nature 437, 426–431 (2005).
Bass, M. D. et al. Syndecan-4-dependent Rac1 regulation determines directional migration in response to the extracellular matrix. J. Cell Biol. 177, 527–538 (2007).
Lachowski, D. et al. Substrate rigidity controls activation and durotaxis in pancreatic stellate cells. Sci. Rep. 7, 2506 (2017).
Várnai, P. & Balla, T. Visualization of phosphoinositides that bind pleckstrin homology domains: calcium- and agonist-induced dynamic changes and relationship to Myo-[3H]inositol-labeled phosphoinositide pools. J. Cell Biol. 143, 501–510 (1998).
Edlund, M., Lotano, M. A. & Otey, C. A. Dynamics of α-actinin in focal adhesions and stress fibers visualized with α-actinin-green fluorescent protein. Cell Motil. 48, 190–200 (2001).
Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).
Jo, S., Kim, T., Iyer, V. G. & Im, W. CHARMM-GUI: a web-based graphical user interface for CHARMM. J. Comput. Chem. 29, 1859–1865 (2008).
Javanainen, M. Universal method for embedding proteins into complex lipid bilayers for molecular dynamics simulations. J. Chem. Theory Comput. 10, 2577–2582 (2014).
Van Der Spoel, D. et al. GROMACS: fast, flexible and free. J. Comput. Chem. 26, 1701–1718 (2005).
Kaminski, G. A., Friesner, R. A., Tirado-Rives, J. & Jorgensen, W. L. Evaluation and reparametrization of the OPLS-AA force field for proteins via comparison with accurate quantum chemical calculations on peptides. J. Phys. Chem. B 105, 6474–6487 (2001).
Jorgensen, W. L. & Madura, J. D. Quantum and statistical mechanical studies of liquids. 25. Solvation and conformation of methanol in water. J. Am. Chem. Soc. 105, 1407–1413 (1983).
Berendsen, H. J. C., Postma, J. P. M., van Gunsteren, W. F., DiNola, A. & Haak, J. R. Molecular-dynamics with coupling to an external bath. J. Chem. Phys. 81, 3684–3690 (1984).
Hoover, W. G. Canonical dynamics: equilibrium phase-space distributions. Phys. Rev. A 31, 1695–1697 (1985).
Nosé, S. A unified formulation of the constant temperature molecular dynamics methods. J. Chem. Phys. 81, 511–519 (1984).
Parrinello, M. & Rahman, A. Polymorphic transitions in single-crystals—a new molecular-dynamics method. J. Appl. Phys. 52, 7182–7190 (1981).
This work was supported by the European Research Council (ERC grant no. 282051), the Biotechnology and Biological Sciences Research Council (BBSRC grant no. BB/N018532/1) and the Academy of Finland (grant no. 290506). V.V.M. was supported by an EDUFI (former CIMO) postdoctoral fellowship and Academy of Finland funding for Postdoctoral Researcher (grant no. 323021). We thank M. Morgan (University of Liverpool) for providing MEF cell lines, J. Couchman (University of Copenhagen) for providing syndecan-4 cytoplasmic truncation plasmids (C2 and V domains), J. Qin (Cleveland Clinic) for the kindlin-2-GFP plasmids, C. Wu (University of Pittsburgh) for the kindlin-2 K390A plasmid and F. Di Maggio for help in implementing the initial work with PSCs. We acknowledge CSC–IT Center for Science, Finland for computational resources. We are also grateful to all CMBL members for help and advice throughout this work.
The authors declare no competing interests.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Chronopoulos, A., Thorpe, S.D., Cortes, E. et al. Syndecan-4 tunes cell mechanics by activating the kindlin-integrin-RhoA pathway. Nat. Mater. (2020). https://doi.org/10.1038/s41563-019-0567-1