Optogenetic control of RhoA reveals zyxin-mediated elasticity of stress fibres

Patrick W. Oakes; Elizabeth Wagner; Christoph A. Brand; Dimitri Probst; Marco Linke; Ulrich S. Schwarz; Michael Glotzer; Margaret L. Gardel

doi:10.1038/ncomms15817

journal article Open Access Jun 12, 2017

Optogenetic control of RhoA reveals zyxin-mediated elasticity of stress fibres

Patrick W. Oakes

Elizabeth Wagner Christoph A. Brand Dimitri Probst Marco Linke Ulrich S. Schwarz

Michael Glotzer

Margaret L. Gardel

Nature Communications Vol. 8 No. 1 · Springer Science and Business Media LLC

View at Publisher Save 10.1038/ncomms15817

Abstract

Abstract
Cytoskeletal mechanics regulates cell morphodynamics and many physiological processes. While contractility is known to be largely RhoA-dependent, the process by which localized biochemical signals are translated into cell-level responses is poorly understood. Here we combine optogenetic control of RhoA, live-cell imaging and traction force microscopy to investigate the dynamics of actomyosin-based force generation. Local activation of RhoA not only stimulates local recruitment of actin and myosin but also increased traction forces that rapidly propagate across the cell via stress fibres and drive increased actin flow. Surprisingly, this flow reverses direction when local RhoA activation stops. We identify zyxin as a regulator of stress fibre mechanics, as stress fibres are fluid-like without flow reversal in its absence. Using a physical model, we demonstrate that stress fibres behave elastic-like, even at timescales exceeding turnover of constituent proteins. Such molecular control of actin mechanics likely plays critical roles in regulating morphodynamic events.

Topics

No keywords indexed for this article. Browse by subject →

References

76

[1]

Force Generation, Transmission, and Integration during Cell and Tissue Morphogenesis

Thomas Lecuit, Pierre-François Lenne, Edwin Munro

Annual Review of Cell and Developmental Biology 2011 10.1146/annurev-cellbio-100109-104027

[2]

Paluch, E. & Heisenberg, C.-P. Biology and physics of cell shape changes in development. Curr. Biol. 19, R790–R799 (2009). 10.1016/j.cub.2009.07.029

[3]

Iskratsch, T., Wolfenson, H. & Sheetz, M. P. Appreciating force and shape—the rise of mechanotransduction in cell biology. Nat. Rev. Mol. Cell Biol. 15, 825–833 (2014). 10.1038/nrm3903

[4]

Murrell, M., Oakes, P. W., Lenz, M. & Gardel, M. L. Forcing cells into shape: the mechanics of actomyosin contractility. Nat. Rev. Mol. Cell Biol. 16, 486–498 (2015). 10.1038/nrm4012

[5]

Martin, A. C., Gelbart, M., Fernandez-Gonzalez, R., Kaschube, M. & Wieschaus, E. F. Integration of contractile forces during tissue invagination. J. Cell Biol. 188, 735–749 (2010). 10.1083/jcb.200910099

[6]

Wagner, E. & Glotzer, M. Local RhoA activation induces cytokinetic furrows independent of spindle position and cell cycle stage. J. Cell Biol. 213, 641–649 (2016). 10.1083/jcb.201603025

[7]

Melvin, A. T., Welf, E. S., Wang, Y., Irvine, D. J. & Haugh, J. M. In chemotaxing fibroblasts, both high-fidelity and weakly biased cell movements track the localization of PI3K signaling. Biophys. J. 100, 1893–1901 (2011). 10.1016/j.bpj.2011.02.047

[8]

Plotnikov, S. V. & Waterman, C. M. Guiding cell migration by tugging. Curr. Opin. Cell Biol. 25, 619–626 (2013). 10.1016/j.ceb.2013.06.003

[9]

Cell Shape, Cytoskeletal Tension, and RhoA Regulate Stem Cell Lineage Commitment

Rowena McBeath, Dana M Pirone, Celeste M Nelson et al.

Developmental Cell 2004 10.1016/s1534-5807(04)00075-9

[10]

Wang, A. et al. Nonmuscle myosin II isoform and domain specificity during early mouse development. Proc. Natl Acad. Sci. USA 107, 14645–14650 (2010). 10.1073/pnas.1004023107

[11]

Müller, T. et al. Non-muscle myosin IIA is required for the development of the zebrafish glomerulus. Kidney Int. 80, 1055–1063 (2011). 10.1038/ki.2011.256

[12]

Schwarz, U. S. & Gardel, M. L. United we stand - integrating the actin cytoskeleton and cell-matrix adhesions in cellular mechanotransduction. J. Cell Sci. 125, 3051–3060 (2012). 10.1242/jcs.093716

[13]

Actin Dynamics, Architecture, and Mechanics in Cell Motility

Laurent Blanchoin, Rajaa Boujemaa-Paterski, Cécile Sykes et al.

Physiological Reviews 2014 10.1152/physrev.00018.2013

[14]

Cell mechanics and the cytoskeleton

Daniel A. Fletcher, R. Dyche Mullins

Nature 2010 10.1038/nature08908

[15]

Hotulainen, P. & Lappalainen, P. Stress fibers are generated by two distinct actin assembly mechanisms in motile cells. J. Cell Biol. 173, 383–394 (2006). 10.1083/jcb.200511093

[16]

Devreotes, P. & Horwitz, A. R. Signaling networks that regulate cell migration. Cold Spring Harb. Perspect. Biol. 7, a005959 (2015). 10.1101/cshperspect.a005959

[17]

Gardel, M. L., Schneider, I. C., Aratyn-Schaus, Y. & Waterman, C. M. Mechanical integration of actin and adhesion dynamics in cell migration. Annu. Rev. Cell Dev. Biol. 26, 315–333 (2010). 10.1146/annurev.cellbio.011209.122036

[18]

Hoffman, B. D. & Crocker, J. C. Cell mechanics: dissecting the physical responses of cells to force. Annu. Rev. Biomed. Eng. 11, 259–288 (2009). 10.1146/annurev.bioeng.10.061807.160511

[19]

Mechanics of the F-actin cytoskeleton

Jonathan Stricker, Tobias Falzone, Margaret L. Gardel

Journal of Biomechanics 2010 10.1016/j.jbiomech.2009.09.003

[20]

Zhu, C., Bao, G. & Wang, N. Cell mechanics: mechanical response, cell adhesion, and molecular deformation. Annu. Rev. Biomed. Eng. 2, 189–226 (2000). 10.1146/annurev.bioeng.2.1.189

[21]

Active gel physics

J. Prost, F. Jülicher, J-F. Joanny

Nature Physics 2015 10.1038/nphys3224

[22]

Schwarz, U. S. & Safran, S. A. Physics of adherent cells. Rev. Mod. Phys. 85, 1327 (2013). 10.1103/revmodphys.85.1327

[23]

Broedersz, C. P. & Mackintosh, F. C. Modeling semiflexible polymer networks. Rev. Mod. Phys. 86, 995 (2014). 10.1103/revmodphys.86.995

[24]

Fritzsche, M., Lewalle, A., Duke, T., Kruse, K. & Charras, G. Analysis of turnover dynamics of the submembranous actin cortex. Mol. Biol. Cell 24, 757–767 (2013). 10.1091/mbc.e12-06-0485

[25]

Saha, A. et al. Determining physical properties of the cell cortex. Biophys. J. 110, 1421–1429 (2016). 10.1016/j.bpj.2016.02.013

[26]

Hu, S. et al. Long-range self-organization of cytoskeletal myosin II filament stacks. Nat. Cell Biol. 19, 133–141 (2017). 10.1038/ncb3466

[27]

Ridley, A. J. & Hall, A. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell 70, 389–399 (1992). 10.1016/0092-8674(92)90163-7

[28]

Chrzanowska-Wodnicka, M. & Burridge, K. Rho-stimulated contractility drives the formation of stress fibers and focal adhesions. J. Cell Biol. 133, 1403–1415 (1996). 10.1083/jcb.133.6.1403

[29]

Coordination of Rho GTPase activities during cell protrusion

Matthias Machacek, Louis Hodgson, Christopher Welch et al.

Nature 2009 10.1038/nature08242

[30]

Pertz, O., Hodgson, L., Klemke, R. L. & Hahn, K. M. Spatiotemporal dynamics of RhoA activity in migrating cells. Nature 440, 1069–1072 (2006). 10.1038/nature04665

[31]

A genetically encoded photoactivatable Rac controls the motility of living cells

Yi I. Wu, Daniel Frey, Oana I. Lungu et al.

Nature 2009 10.1038/nature08241

[32]

Weitzman, M. & Hahn, K. M. Optogenetic approaches to cell migration and beyond. Curr. Opin. Cell Biol. 30, 112–120 (2014). 10.1016/j.ceb.2014.08.004

[33]

Wu, J. et al. Actomyosin pulls to advance the nucleus in a migrating tissue cell. Biophys. J. 106, 7–15 (2014). 10.1016/j.bpj.2013.11.4489

[34]

Valon, L., Marín-Llauradó, A., Wyatt, T., Charras, G. & Trepat, X. Optogenetic control of cellular forces and mechanotransduction. Nat. Commun. 8, 14396 (2017). 10.1038/ncomms14396

[35]

TULIPs: tunable, light-controlled interacting protein tags for cell biology

Devin Strickland, Yuan Lin, Elizabeth Wagner et al.

Nature Methods 2012 10.1038/nmeth.1904

[36]

The Rho GEFs LARG and GEF-H1 regulate the mechanical response to force on integrins

Christophe Guilluy, Vinay Swaminathan, Rafael Garcia-Mata et al.

Nature Cell Biology 2011 10.1038/ncb2254

[37]

Jaiswal, M. et al. Mechanistic insights into specificity, activity, and regulatory elements of the regulator of G-protein signaling (RGS)-containing Rho-specific guanine nucleotide exchange factors (GEFs) p115, PDZ-RhoGEF (PRG), and leukemia-associated RhoGEF (LARG). J. Biol. Chem. 286, 18202–18212 (2011). 10.1074/jbc.m111.226431

[38]

Rizvi, S. A. et al. Identification and characterization of a small molecule inhibitor of formin-mediated actin assembly. Chem. Biol. 16, 1158–1168 (2009). 10.1016/j.chembiol.2009.10.006

[39]

Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates

Nathalie Q. Balaban, Ulrich S. Schwarz, Daniel Riveline et al.

Nature Cell Biology 2001 10.1038/35074532

[40]

Sabass, B., Gardel, M., Waterman, C. M. & Schwarz, U. S. High resolution traction force microscopy based on experimental and computational advances. Biophys. J. 94, 207–220 (2008). 10.1529/biophysj.107.113670

[41]

Soiné, J. R. D. et al. Model-based traction force microscopy reveals differential tension in cellular actin bundles. PLoS Comput. Biol. 11, e1004076 (2015). 10.1371/journal.pcbi.1004076

[42]

Stricker, J., Aratyn-Schaus, Y., Oakes, P. W. & Gardel, M. L. Spatiotemporal constraints on the force-dependent growth of focal adhesions. Biophys. J. 100, 2883–2893 (2011). 10.1016/j.bpj.2011.05.023

[43]

Tension is required but not sufficient for focal adhesion maturation without a stress fiber template

Patrick W. Oakes, Yvonne Beckham, Jonathan Stricker et al.

Journal of Cell Biology 2012 10.1083/jcb.201107042

[44]

Geometry Regulates Traction Stresses in Adherent Cells

Patrick W. Oakes, Shiladitya Banerjee, M. Cristina Marchetti et al.

Biophysical Journal 2014 10.1016/j.bpj.2014.06.045

[45]

Brown, R. A., Prajapati, R., McGrouther, D. A., Yannas, I. V. & Eastwood, M. Tensional homeostasis in dermal fibroblasts: mechanical responses to mechanical loading in three-dimensional substrates. J. Cell Physiol. 175, 323–332 (1998). 10.1002/(sici)1097-4652(199806)175:3<323::aid-jcp10>3.0.co;2-6

[46]

Tensegrity I. Cell structure and hierarchical systems biology

Donald E. Ingber

Journal of Cell Science 2003 10.1242/jcs.00359

[47]

Webster, K. D., Ng, W. P. & Fletcher, D. A. Tensional homeostasis in single fibroblasts. Biophys. J. 107, 146–155 (2014). 10.1016/j.bpj.2014.04.051

[48]

Turlier, H., Audoly, B., Prost, J. & Joanny, J.-F. Furrow constriction in animal cell cytokinesis. Biophys. J. 106, 114–123 (2014). 10.1016/j.bpj.2013.11.014

[49]

High Accuracy Optical Flow Estimation Based on a Theory for Warping

Thomas Brox, Andrés Bruhn, Nils Papenberg et al.

Lecture Notes in Computer Science 10.1007/978-3-540-24673-2_3

[50]

Sánchez, J., Monzón, N. & Salgado, A. Robust optical flow estimation. Image Process. On Line 3, 252–270 (2013). 10.5201/ipol.2013.21

Showing 50 of 76 references

Metrics

174

Citations

76

References

Details

Published: Jun 12, 2017
Vol/Issue: 8(1)
License: View

Authors

P

Patrick W. Oakes

Institute for Biophysical Dynamics 1 , 2 , and 3; Department of Physics 1 , 2 , and 3; The James Franck Institute, University of Chicago, Chicago, IL 60637 1 , 2 , and 3; Institute for Biophysical Dynamics 1 , 2 , and 3; Department of Physics 1 , 2 , and 3; The James Franck Institute, University of Chicago, Chicago, IL 60637 1 , 2 , and 3; Institute for Biophysical Dynamics 1 , 2 , and 3; Department of Physics 1 , 2 , and 3; The James Franck Institute, University of Chicago, Chicago, IL 60637 1 , 2 , and 3

Institute for Biophysical Dynamics 1 , 2 , and 3; Department of Physics 1 , 2 , and 3; The James Franck Institute, University of Chicago, Chicago, IL 60637 1 , 2 , and 3; Institute for Biophysical Dynamics 1 , 2 , and 3; Department of Physics 1 , 2 , and 3; The James Franck Institute, University of Chicago, Chicago, IL 60637 1 , 2 , and 3; Institute for Biophysical Dynamics 1 , 2 , and 3; Department of Physics 1 , 2 , and 3; The James Franck Institute, University of Chicago, Chicago, IL 60637 1 , 2 , and 3

Cite This Article

Patrick W. Oakes, Elizabeth Wagner, Christoph A. Brand, et al. (2017). Optogenetic control of RhoA reveals zyxin-mediated elasticity of stress fibres. Nature Communications, 8(1). https://doi.org/10.1038/ncomms15817

Optogenetic control of RhoA reveals zyxin-mediated elasticity of stress fibres

You May Also Like