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Integration of actin dynamics and cell adhesion by a three-dimensional, mechanosensitive molecular clutch

Nature Cell Biology volume 17, pages 955963 (2015) | Download Citation

Abstract

During cell migration, the forces generated in the actin cytoskeleton are transmitted across transmembrane receptors to the extracellular matrix or other cells through a series of mechanosensitive, regulable protein–protein interactions termed the molecular clutch. In integrin-based focal adhesions, the proteins forming this linkage are organized into a conserved three-dimensional nano-architecture. Here we discuss how the physical interactions between the actin cytoskeleton and focal-adhesion-associated molecules mediate force transmission from the molecular clutch to the extracellular matrix.

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References

  1. 1.

    Prespecification and plasticity: shifting mechanisms of cell migration. Curr. Opin. Cell Biol. 16, 14–23 (2004).

  2. 2.

    & The role and regulation of blebs in cell migration. Curr. Opin. Cell Biol. 25, 582–590 (2013).

  3. 3.

    et al. Water permeation drives tumor cell migration in confined microenvironments. Cell 157, 611–623 (2014).

  4. 4.

    et al. Adaptive force transmission in amoeboid cell migration. Nat. Cell Biol. 11, 1438–1443 (2009).

  5. 5.

    , & Generation of compartmentalized pressure by a nuclear piston governs cell motility in a 3D matrix. Science 345, 1062–1065 (2014).

  6. 6.

    & Actin, a central player in cell shape and movement. Science 326, 1208–1212 (2009).

  7. 7.

    et al. Confinement and low adhesion induce fast amoeboid migration of slow mesenchymal cells. Cell 160, 659–672 (2015).

  8. 8.

    et al. Cortical contractility triggers a stochastic switch to fast amoeboid cell motility. Cell 160, 673–685 (2015).

  9. 9.

    , & Random versus directionally persistent cell migration. Nat. Rev. Mol. Cell Biol. 10, 538–549 (2009).

  10. 10.

    Exchange of actin subunits at the leading edge of living fibroblasts: possible role of treadmilling. J. Cell Biol. 101, 597–602 (1985).

  11. 11.

    & Actin microfilament dynamics in locomoting cells. Nature 352, 126–131 (1991).

  12. 12.

    & Cell motility driven by actin polymerization. Biophys. J. 71, 3030–3045 (1996).

  13. 13.

    & Growth cone advance is inversely proportional to retrograde F-actin flow. Neuron 14, 763–771 (1995).

  14. 14.

    & Cytoskeletal dynamics and nerve growth. Neuron 1, 761–772 (1988).

  15. 15.

    , & Regulation of RhoA activity by adhesion molecules and mechanotransduction. Curr. Mol. Med. 14, 199–208 (2014).

  16. 16.

    , & Integrin modulation and signaling in leukocyte adhesion and migration. Immunol. Rev. 218, 126–134 (2007).

  17. 17.

    & Cell motility under the microscope: Vorsprung durch Technik. Nat. Rev. Mol. Cell Biol. 5, 667–672 (2004).

  18. 18.

    The bases of the locomotory behaviour of fibroblasts. Exp. Cell Res. 8, 188–198 (1961).

  19. 19.

    , & The locomotion of fibroblasts in culture: I. Movements of the leading edge. Exp. Cell Res. 59, 393–398 (1970).

  20. 20.

    , & The locomotion of fibroblasts in culture: III. Movements of particles on the dorsal surface of the leading lamella. Exp. Cell Res. 62, 389–398 (1970).

  21. 21.

    & Centripetal transport of attached particles on both surfaces of moving fibroblasts. Exp. Cell Res. 73, 519–523 (1972).

  22. 22.

    Cell surface movements related to cell locomotion. Ciba Found. Symp. 14, 3–26 (1973).

  23. 23.

    , & The locomotion of fibroblasts in culture: IV. Electron microscopy of the leading lamella. Exp. Cell Res. 67, 359–367 (1971).

  24. 24.

    , & Polarity of actin at the leading edge of cultured cells. Nature 272, 638–639 (1978).

  25. 25.

    Actions of cytochalasins on the organization of actin filaments and microtubules in a neuronal growth cone. J. Cell Biol. 107, 1505–1516 (1988).

  26. 26.

    , , & Principles of locomotion for simple-shaped cells. Nature 362, 167–171 (1993).

  27. 27.

    et al. Two components of actin-based retrograde flow in sea urchin coelomocytes. Mol. Biol. Cell 10, 4075–4090 (1999).

  28. 28.

    , , , & Two distinct actin networks drive the protrusion of migrating cells. Science 305, 1782–1786 (2004).

  29. 29.

    Molecular mechanism of actin-dependent retrograde flow in lamellipodia of motile cells. Front. Biosci. 2, 260–70 (1997).

  30. 30.

    , , & Actin polymerization induces a shape change in actin-containing vesicles. Proc. Natl Acad. Sci. USA 86, 5773–5777 (1989).

  31. 31.

    & In silico reconstitution of Listeria propulsion exhibits nano-saltation. PLoS Biol. 2, e412 (2004).

  32. 32.

    & Spatial and temporal relationships between actin-filament nucleation, capping, and disassembly. Curr. Biol. 17, 395–406 (2007).

  33. 33.

    & Stress fibers are generated by two distinct actin assembly mechanisms in motile cells. J. Cell Biol. 173, 383–394 (2006).

  34. 34.

    V, , & Assembling an actin cytoskeleton for cell attachment and movement. Biochim. Biophys. Acta 1404, 271–281 (1998).

  35. 35.

    , , & Assembly of non-contractile dorsal stress fibers requires α-actinin-1 and Rac1 in migrating and spreading cells. J. Cell Sci. 126, 263–73 (2013).

  36. 36.

    , & Myosin II filament assemblies in the active lamella of fibroblasts: their morphogenesis and role in the formation of actin filament bundles. J. Cell Biol. 131, 989–1002 (1995).

  37. 37.

    et al. Myosin II contributes to cell-scale actin network treadmilling through network disassembly. Nature 465, 373–377 (2010).

  38. 38.

    & Actomyosin-based retrograde flow of microtubules in the lamella of migrating epithelial cells influences microtubule dynamic instability and turnover and is associated with microtubule breakage and treadmilling. J. Cell Biol. 139, 417–434 (1997).

  39. 39.

    et al. Traction stress in focal adhesions correlates biphasically with actin retrograde flow speed. J. Cell Biol. 183, 999–1005 (2008).

  40. 40.

    , & Myosin II functions in actin-bundle turnover in neuronal growth cones. Nat. Cell Biol. 8, 215–226 (2006).

  41. 41.

    , & Slipping or gripping? Fluorescent speckle microscopy in fish keratocytes reveals two different mechanisms for generating a retrograde flow of actin. Mol. Biol. Cell 16, 507–518 (2005).

  42. 42.

    , , , & Tracking retrograde flow in keratocytes: news from the front. Mol. Biol. Cell 16, 1223–1231 (2005).

  43. 43.

    et al. Spatio-temporal analysis of eukaryotic cell motility by improved force cytometry. Proc. Natl Acad. Sci. USA 104, 13343–13348 (2007).

  44. 44.

    & Relationships between fibronectin (LETS protein) and actin. Cell 15, 875–886 (1978).

  45. 45.

    The fibronexus: a transmembrane association of fibronectin-containing fibers and bundles of 5 nm microfilaments in hamster and human fibroblasts. Cell 16, 675–685 (1979).

  46. 46.

    Association of fibronectin and vinculin with focal contacts and stress fibers in stationary hamster fibroblasts. J. Cell Biol. 92, 398–408 (1982).

  47. 47.

    et al. Structure of integrin, a glycoprotein involved in the transmembrane linkage between fibronectin and actin. Cell 46, 271–282 (1986).

  48. 48.

    Integrins: bidirectional, allosteric signaling machines. Cell 110, 673–687 (2002).

  49. 49.

    , & C-terminal opening mimics “inside-out” activation of integrin α5β1. Nat. Struct. Biol. 8, 412–416 (2001).

  50. 50.

    , , & Global conformational rearrangements in integrin extracellular domains in outside-in and inside-out signaling. Cell 110, 599–611 (2002).

  51. 51.

    et al. The talin head domain binds to integrin subunit cytoplasmic tails and regulates integrin activation. J. Biol. Chem. 274, 28071–28074 (1999).

  52. 52.

    , & An interaction between α-actinin and the β1 integrin subunit in vitro. J. Cell Biol. 111, 721–729 (1990).

  53. 53.

    , & Kindlin-1 and -2 directly bind the C-terminal region of β integrin cytoplasmic tails and exert integrin-specific activation effects. J. Biol. Chem. 284, 11485–11497 (2009).

  54. 54.

    et al. Integrin β cytoplasmic domain interactions with phosphotyrosine-binding domains: a structural prototype for diversity in integrin signaling. Proc. Natl Acad. Sci. USA 100, 2272–2277 (2003).

  55. 55.

    & Mechanisms that regulate adaptor binding to β-integrin cytoplasmic tails. J. Cell Sci. 122, 187–198 (2009).

  56. 56.

    , & Integrin cytoplasmic tail interactions. Biochemistry 53, 810–820 (2014).

  57. 57.

    , , , & Functional atlas of the integrin adhesome. Nat. Cell Biol. 9, 858–867 (2007).

  58. 58.

    , , & Analysis of the myosin-II-responsive focal adhesion proteome reveals a role for β-Pix in negative regulation of focal adhesion maturation. Nat. Cell Biol. 13, 383–393 (2011).

  59. 59.

    , , , & Proteomic analysis of integrin adhesion complexes. Sci. Signal. 4, 2 (2011).

  60. 60.

    , , & Quantitative proteomics of the integrin adhesome show a myosin II-dependent recruitment of LIM domain proteins. EMBO Rep. 12, 259–266 (2011).

  61. 61.

    , , & Early molecular events in the assembly of matrix adhesions at the leading edge of migrating cells. J. Cell Sci. 116, 4605–4613 (2003).

  62. 62.

    et al. Actin and α-actinin orchestrate the assembly and maturation of nascent adhesions in a myosin II motor-independent manner. Nat. Cell Biol. 10, 1039–1050 (2008).

  63. 63.

    et al. FAK promotes recruitment of talin to nascent adhesions to control cell motility. J. Cell Biol. 196, 223–232 (2012).

  64. 64.

    , , & Tension is required but not sufficient for focal adhesion maturation without a stress fiber template. J. Cell Biol. 196, 363–374 (2012).

  65. 65.

    et al. Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes. Proc. Natl Acad. Sci. USA 104, 20308–20313 (2007).

  66. 66.

    et al. Interferometric fluorescent super-resolution microscopy resolves 3D cellular ultrastructure. Proc. Natl Acad. Sci. USA 106, 3125–3130 (2009).

  67. 67.

    et al. Nanoscale architecture of integrin-based cell adhesions. Nature 468, 580–584 (2010).

  68. 68.

    et al. Scanning angle interference microscopy reveals cell dynamics at the nanoscale. Nat. Methods 9, 825–827 (2012).

  69. 69.

    et al. Molecular mechanism of vinculin activation and nanoscale spatial organization in focal adhesions. Nat. Cell Biol. 17, 880–892 (2015).

  70. 70.

    Biochemical and structural properties of the integrin-associated cytoskeletal protein talin. Annu. Rev. Biophys. 38, 235–254 (2009).

  71. 71.

    , , , & Interaction of plasma membrane fibronectin receptor with talin — a transmembrane linkage. Nature 320, 531–533 (1986).

  72. 72.

    et al. Examining F-actin interaction with intact talin and talin head and tail fragment using static and dynamic light scattering. Eur. J. Biochem. 250, 447–450 (1997).

  73. 73.

    et al. A role for the juxtamembrane cytoplasm in the molecular dynamics of focal adhesions. PLoS One 4, e4304 (2009).

  74. 74.

    et al. Molecular diversity of cell-matrix adhesions. J. Cell Sci. 112, 1655–1669 (1999).

  75. 75.

    , , & A paxillin tyrosine phosphorylation switch regulates the assembly and form of cell-matrix adhesions. J. Cell Sci. 120, 137–148 (2007).

  76. 76.

    et al. Mechanotransduction in vivo by repeated talin stretch-relaxation events depends upon vinculin. PLoS Biol. 9, e1001223 (2011).

  77. 77.

    et al. Archipelago architecture of the focal adhesion: membrane molecules freely enter and exit from the focal adhesion zone. Cytoskeleton 69, 380–392 (2012).

  78. 78.

    et al. Integrins β1 and β3 exhibit distinct dynamic nanoscale organizations inside focal adhesions. Nat. Cell Biol. 14, 1057–1067 (2012).

  79. 79.

    , , & Force-dependent integrin-cytoskeleton linkage formation requires downregulation of focal complex dynamics by Shp2. EMBO J. 22, 5023–5035 (2003).

  80. 80.

    , , & Investigating complexity of protein-protein interactions in focal adhesions. Biochem. Biophys. Res. Commun. 369, 929–934 (2008).

  81. 81.

    et al. Differential effect of actomyosin relaxation on the dynamic properties of focal adhesion proteins. PLoS One 8, e73549 (2013).

  82. 82.

    et al. Integrin-associated complexes form hierarchically with variable stoichiometry in ascent adhesions. Curr. Biol. 24, 1845–1853 (2014).

  83. 83.

    , , , & Symmetric exchange of multi-protein building blocks between stationary focal adhesions and the cytosol. eLife 3, e02257 (2014).

  84. 84.

    , , , & Role of focal adhesions and mechanical stresses in the formation and progression of the lamellipodium-lamellum interface [corrected]. Biophys. J. 97, 1254–1264 (2009).

  85. 85.

    et al. Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates. Nat. Cell Biol. 3, 466–472 (2001).

  86. 86.

    et al. Comparative dynamics of retrograde actin flow and focal adhesions: formation of nascent adhesions triggers transition from fast to slow flow. PLoS One 3, e3234 (2008).

  87. 87.

    & Spatiotemporal feedback between actomyosin and focal-adhesion systems optimizes rapid cell migration. Cell 125, 1361–1374 (2006).

  88. 88.

    & Rho-stimulated contractility drives the formation of stress fibers and focal adhesions. J. Cell Biol. 133, 1403–1415 (1996).

  89. 89.

    et al. FAK-Src signalling through paxillin, ERK and MLCK regulates adhesion disassembly. Nat. Cell Biol. 6, 154–161 (2004).

  90. 90.

    et al. Structure of a complete integrin ectodomain in a physiologic resting state and activation and deactivation by applied forces. Mol. Cell 32, 849–861 (2008).

  91. 91.

    & An interaction between integrin and the talin FERM domain mediates integrin activation but not linkage to the cytoskeleton. Nat. Cell Biol. 8, 601–606 (2006).

  92. 92.

    et al. Integrin cytoplasmic domains mediate inside-out signal transduction. J. Cell Biol. 124, 1047–1059 (1994).

  93. 93.

    et al. The structure of an integrin/talin complex reveals the basis of inside-out signal transduction. EMBO J. 28, 3623–3632 (2009).

  94. 94.

    , & Modulation of cell migration by integrin-mediated cytoskeletal linkages and ligand-binding affinity. J Cell Biol 134, 1551–1562 (1996).

  95. 95.

    , , , & FRET detection of cellular α4-integrin conformational activation. Biophys. J. 85, 3951–3962 (2003).

  96. 96.

    , V & Bidirectional transmembrane signaling by cytoplasmic domain separation in integrins. Science 301, 1720–1725 (2003).

  97. 97.

    et al. Arp2/3 is critical for lamellipodia and response to extracellular matrix cues but is dispensable for chemotaxis. Cell 148, 973–987 (2012).

  98. 98.

    et al. Arp2/3 inhibition induces amoeboid-like protrusions in MCF10A epithelial cells by reduced cytoskeletal-membrane coupling and focal adhesion assembly. PLoS One 9, e100943 (2014).

  99. 99.

    , & Recruitment of the Arp2/3 complex to vinculin: coupling membrane protrusion to matrix adhesion. J. Cell Biol. 159, 881–891 (2002).

  100. 100.

    et al. Focal adhesion kinase controls actin assembly via a FERM-mediated interaction with the Arp2/3 complex. Nat. Cell Biol. 9, 1046–1056 (2007).

  101. 101.

    , , & Differential dynamics of α5 integrin, paxillin, and α-actinin during formation and disassembly of adhesions in migrating cells. J. Cell Biol. 153, 1427–1440 (2001).

  102. 102.

    , , , & Regulation of protrusion, adhesion dynamics, and polarity by myosins IIA and IIB in migrating cells. J. Cell Biol. 176, 573–580 (2007).

  103. 103.

    et al. Vinculin-actin interaction couples actin retrograde flow to focal adhesions, but is dispensable for focal adhesion growth. J. Cell Biol. 202, 163–177 (2013).

  104. 104.

    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).

  105. 105.

    , & The relationship between force and focal complex development. J. Cell Biol. 159, 695–705 (2002).

  106. 106.

    et al. Caldesmon inhibits nonmuscle cell contractility and interferes with the formation of focal adhesions. Mol. Biol. Cell 10, 3097–3112 (1999).

  107. 107.

    , , & Focal adhesions as mechanosensors: a physical mechanism. Proc. Natl Acad. Sci. USA 102, 12383–12388 (2005).

  108. 108.

    , & Mechanotransduction across the cell surface and through the cytoskeleton. Science 260, 1124–1127 (1993).

  109. 109.

    , & Extracellular matrix rigidity causes strengthening of integrin-cytoskeleton linkages. Cell 88, 39–48 (1997).

  110. 110.

    , , & mDia2 regulates actin and focal adhesion dynamics and organization in the lamella for efficient epithelial cell migration. J. Cell Sci. 120, 3475–3587 (2007).

  111. 111.

    V, , & Force fluctuations within focal adhesions mediate ECM-rigidity sensing to guide directed cell migration. Cell 151, 1513–1527 (2012).

  112. 112.

    et al. Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics. Nature 466, 263–266 (2010).

  113. 113.

    , , & Tension sensing nanoparticles for mechano-imaging at the living/nonliving interface. J. Am. Chem. Soc. 135, 5320–5323 (2013).

  114. 114.

    , , & Molecular tension sensors report forces generated by single integrin molecules in living cells. Nano Lett. 13, 3985–3989 (2013).

  115. 115.

    & Defining single molecular forces required to activate integrin and notch signaling. Science 340, 991–994 (2013).

  116. 116.

    , , , & Integrin-generated forces lead to streptavidin-biotin unbinding in cellular adhesions. Biophys. J. 106, 1436–1446 (2014).

  117. 117.

    et al. Force sensing by mechanical extension of the Src family kinase substrate p130Cas. Cell 127, 1015–1026 (2006).

  118. 118.

    , , , & Demonstration of catch bonds between an integrin and its ligand. J. Cell Biol. 185, 1275–1284 (2009).

  119. 119.

    , , , & Two-piconewton slip bond between fibronectin and the cytoskeleton depends on talin. Nature 424, 334–337 (2003).

  120. 120.

    et al. Stretching single talin rod molecules activates vinculin binding. Science 323, 638–641 (2009).

  121. 121.

    et al. Mechanical activation of vinculin binding to talin locks talin in an unfolded conformation. Sci. Rep. 4, 4610 (2014).

  122. 122.

    & Processive capping by formin suggests a force-driven mechanism of actin polymerization. J. Cell Biol. 167, 1011–1017 (2004).

  123. 123.

    , & Formin mDia1 senses and generates mechanical forces on actin filaments. Nat. Commun. 4, 1883 (2013).

  124. 124.

    , , , & Differential transmission of actin motion within focal adhesions. Science 315, 111–115 (2007).

  125. 125.

    , , & High resolution traction force microscopy based on experimental and computational advances. Biophys. J. 94, 207–220 (2008).

  126. 126.

    et al. Probing the integrin-actin linkage using high-resolution protein velocity mapping. J. Cell Sci. 119, 5204–5214 (2006).

  127. 127.

    & Traction dynamics of filopodia on compliant substrates. Science 322, 1687–1691 (2008).

  128. 128.

    , & Physical model for self-organization of actin cytoskeleton and adhesion complexes at the cell front. Biophys. J. 102, 1746–1756 (2012).

  129. 129.

    , , , & Talin1 is critical for force-dependent reinforcement of initial integrin-cytoskeleton bonds but not tyrosine kinase activation. J. Cell Biol. 163, 409–419 (2003).

  130. 130.

    et al. Talin depletion reveals independence of initial cell spreading from integrin activation and traction. Nat. Cell Biol. 10, 1062–1068 (2008).

  131. 131.

    , & Coincidence of actin filaments and talin is required to activate vinculin. J. Biol. Chem. 281, 40389–40398 (2006).

  132. 132.

    et al. How vinculin regulates force transmission. Proc. Natl Acad. Sci. USA 110, 9788–9793 (2013).

  133. 133.

    , , & Head/tail interaction of vinculin influences cell mechanical behavior. Biochem. Biophys. Res. Commun. 406, 85–88 (2011).

  134. 134.

    et al. Integrin-dependent force transmission to the extracellular matrix by α-actinin triggers adhesion maturation. Proc. Natl Acad. Sci. USA 110, 1361–1370 (2013).

  135. 135.

    et al. Periodic lamellipodial contractions correlate with rearward actin waves. Cell 116, 431–443 (2004).

  136. 136.

    , & Dynamic myosin phosphorylation regulates contractile pulses and tissue integrity during epithelial morphogenesis. J. Cell Biol. 206, 435–450 (2014).

  137. 137.

    et al. Lamellipodial actin mechanically links myosin activity with adhesion-site formation. Cell 128, 561–575 (2007).

  138. 138.

    Mechanisms of leukocyte transendothelial migration. Annu. Rev. Pathol. 6, 323–344 (2011).

  139. 139.

    , , & Evidence of a functional role for interaction between ICAM-1 and nonmuscle α-actinins in leukocyte diapedesis. J. Immunol. 177, 4113–4121 (2006).

  140. 140.

    et al. Release of cellular tension signals self-restorative ventral lamellipodia to heal barrier micro-wounds. J. Cell Biol. 201, 449–465 (2013).

  141. 141.

    Cell adhesion molecules and actin cytoskeleton at immune synapses and kinapses. Curr. Opin. Cell Biol. 19, 529–533 (2007).

  142. 142.

    , , & Cellular adhesion molecules as targets for bacterial infection. Eur. J. Cell Biol. 85, 235–242 (2006).

  143. 143.

    , , , & Structural mimicry for vinculin activation by IpaA, a virulence factor of Shigella flexneri. EMBO Rep. 7, 794–799 (2006).

  144. 144.

    & Cadherin adhesion and mechanotransduction. Annu. Rev. Cell Dev. Biol. 30, 291–315 (2014).

  145. 145.

    et al. A molecular clutch between the actin flow and N-cadherin adhesions drives growth cone migration. J. Neurosci. 28, 5879–5890 (2008).

  146. 146.

    & Basal-to-apical cadherin flow at cell junctions. Nat. Cell Biol. 9, 92–98 (2007).

  147. 147.

    & Cell movements during vertebrate development: integrated tissue behaviour versus individual cell migration. Curr. Opin. Genet. Dev. 11, 464–469 (2001).

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Acknowledgements

The authors thank Michelle Baird and Michael Davidson (Florida State University) for assistance with figure design and members of the Waterman Lab for helpful discussions. Funding was provided by the Division of Intramural Research, NHLBI (L.B.C. and C.M.W.).

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  1. Lindsay B. Case and Clare M. Waterman are at the Cell Biology and Physiology Center, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, 20892, USA

    • Lindsay B. Case
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Correspondence to Clare M. Waterman.

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