Investigating piconewton forces in cells by FRET-based molecular force microscopy.

The ability of cells to sense and respond to mechanical forces is crucial for a wide range of developmental and pathophysiological processes. The molecular mechanisms underlying cellular mechanotransduction, however, are largely unknown because suitable techniques to measure mechanical forces across individual molecules in cells have been missing. In this article, we highlight advances in the development of molecular force sensing techniques and discuss our recently expanded set of FRET-based tension sensors that allows the analysis of mechanical forces with piconewton sensitivity in cells. In addition, we provide a theoretical framework for the design of additional tension sensor modules with adjusted force sensitivity.

[1]  Albert C. Chen,et al.  Matrix stiffness drives Epithelial-Mesenchymal Transition and tumour metastasis through a TWIST1-G3BP2 mechanotransduction pathway , 2015, Nature Cell Biology.

[2]  Jacob W J Kerssemakers,et al.  Magnetic torque tweezers: measuring torsional stiffness in DNA and RecA-DNA filaments , 2010, Nature Methods.

[3]  Matthias Rief,et al.  Ultrafast folding kinetics and cooperativity of villin headpiece in single-molecule force spectroscopy , 2013, Proceedings of the National Academy of Sciences.

[4]  H. Gaub,et al.  Adhesion forces between individual ligand-receptor pairs. , 1994, Science.

[5]  P. Kollman,et al.  Pathways to a protein folding intermediate observed in a 1-microsecond simulation in aqueous solution. , 1998, Science.

[6]  David R. Liu,et al.  A DNA-based molecular probe for optically reporting cellular traction forces , 2014, Nature Methods.

[7]  E. Michael Ostap,et al.  Myosin I Can Act As a Molecular Force Sensor , 2008, Science.

[8]  M. Krieg,et al.  Tensile forces govern germ-layer organization in zebrafish , 2008, Nature Cell Biology.

[9]  Frank Schnorrer,et al.  Tension and Force-Resistant Attachment Are Essential for Myofibrillogenesis in Drosophila Flight Muscle , 2014, Current Biology.

[10]  Enrico Gratton,et al.  Fluid Shear Stress on Endothelial Cells Modulates Mechanical Tension across VE-Cadherin and PECAM-1 , 2013, Current Biology.

[11]  K. Salaita,et al.  Visualizing mechanical tension across membrane receptors with a fluorescent sensor , 2011, Nature Methods.

[12]  J. Onuchic,et al.  Theory of protein folding: the energy landscape perspective. , 1997, Annual review of physical chemistry.

[13]  Pablo Hernández-Varas,et al.  A plastic relationship between vinculin-mediated tension and adhesion complex area defines adhesion size and lifetime , 2015, Nature communications.

[14]  J. Fredberg,et al.  Collective cell guidance by cooperative intercellular forces , 2010, Nature materials.

[15]  Sami Alom Ruiz,et al.  Mechanical tugging force regulates the size of cell–cell junctions , 2010, Proceedings of the National Academy of Sciences.

[16]  Ching-Wei Chang,et al.  Vinculin tension distributions of individual stress fibers within cell–matrix adhesions , 2013, Journal of Cell Science.

[17]  Beth L. Pruitt,et al.  E-cadherin is under constitutive actomyosin-generated tension that is increased at cell–cell contacts upon externally applied stretch , 2012, Proceedings of the National Academy of Sciences.

[18]  Richard Superfine,et al.  Isolated nuclei adapt to force and reveal a mechanotransduction pathway in the nucleus , 2014, Nature Cell Biology.

[19]  Jens Friedrichs,et al.  Revealing Early Steps of α2β1 Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy , 2007 .

[20]  A. Dunn,et al.  Molecular tension sensors report forces generated by single integrin molecules in living cells. , 2013, Nano letters.

[21]  Taekjip Ha,et al.  Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics , 2010, Nature.

[22]  M. Rief,et al.  Reversible unfolding of individual titin immunoglobulin domains by AFM. , 1997, Science.

[23]  M. Sixt,et al.  Mechanisms of force generation and force transmission during interstitial leukocyte migration , 2010, EMBO reports.

[24]  Andrew S. LaCroix,et al.  Controlling Cell Geometry Affects the Spatial Distribution of Load Across Vinculin , 2015 .

[25]  Gabriel Zoldák,et al.  Force as a single molecule probe of multidimensional protein energy landscapes. , 2013, Current opinion in structural biology.

[26]  P. Hordijk,et al.  Cell-stiffness-induced mechanosignaling – a key driver of leukocyte transendothelial migration , 2015, Journal of Cell Science.

[27]  R. Wells Tissue mechanics and fibrosis. , 2013, Biochimica et biophysica acta.

[28]  K. Salaita,et al.  Lighting Up the Force: Investigating Mechanisms of Mechanotransduction Using Fluorescent Tension Probes , 2015, Molecular and Cellular Biology.

[29]  J. Lammerding,et al.  The cellular mastermind(?)-mechanotransduction and the nucleus. , 2014, Progress in molecular biology and translational science.

[30]  Daniel Choquet,et al.  Extracellular Matrix Rigidity Causes Strengthening of Integrin–Cytoskeleton Linkages , 1997, Cell.

[31]  K. Salaita,et al.  Tension sensing nanoparticles for mechano-imaging at the living/nonliving interface. , 2013, Journal of the American Chemical Society.

[32]  Andrew D. Franck,et al.  The Ndc80 Kinetochore Complex Forms Load-Bearing Attachments to Dynamic Microtubule Tips via Biased Diffusion , 2009, Cell.

[33]  Beat Imhof,et al.  The inner lives of focal adhesions. , 2002, Trends in cell biology.

[34]  Hermann E. Gaub,et al.  Discrete interactions in cell adhesion measured by single-molecule force spectroscopy , 2000, Nature Cell Biology.

[35]  Donald E Ingber,et al.  Mechanical control of tissue and organ development , 2010, Development.

[36]  E. Richter,et al.  Exercise, GLUT4, and skeletal muscle glucose uptake. , 2013, Physiological reviews.

[37]  M. Davidson,et al.  The cancer glycocalyx mechanically primes integrin-mediated growth and survival , 2014, Nature.

[38]  Carsten Grashoff,et al.  How to Measure Molecular Forces in Cells: A Guide to Evaluating Genetically-Encoded FRET-Based Tension Sensors , 2014, Cellular and Molecular Bioengineering.

[39]  M. Dembo,et al.  Stresses at the cell-to-substrate interface during locomotion of fibroblasts. , 1999, Biophysical journal.

[40]  Andrew D. McAinsh,et al.  Molecular control of kinetochore-microtubule dynamics and chromosome oscillations , 2010, Nature Cell Biology.

[41]  A. Bensimon,et al.  The Elasticity of a Single Supercoiled DNA Molecule , 1996, Science.

[42]  A. Schwab,et al.  Endothelial f-actin depolymerization enables leukocyte transmigration , 2011, Analytical and bioanalytical chemistry.

[43]  E. Salmon,et al.  Welcome to a new kind of tension: translating kinetochore mechanics into a wait-anaphase signal , 2010, Journal of Cell Science.

[44]  J. Fredberg,et al.  Mechanical waves during tissue expansion , 2012, Nature Physics.

[45]  Shai Shaham,et al.  FBN-1, a fibrillin-related protein, is required for resistance of the epidermis to mechanical deformation during C. elegans embryogenesis , 2015, eLife.

[46]  Cynthia A. Reinhart-King,et al.  Tensional homeostasis and the malignant phenotype. , 2005, Cancer cell.

[47]  Cheng Zhu,et al.  DNA-based digital tension probes reveal integrin forces during early cell adhesion , 2014, Nature Communications.

[48]  Eric F. Wieschaus,et al.  Integration of contractile forces during tissue invagination , 2010, The Journal of cell biology.

[49]  U. Müller,et al.  Mechanotransduction by Hair Cells: Models, Molecules, and Mechanisms , 2009, Cell.

[50]  Guillermo A. Gomez,et al.  Tension-Sensitive Actin Assembly Supports Contractility at the Epithelial Zonula Adherens , 2014, Current Biology.

[51]  Gaudenz Danuser,et al.  Mechanical Feedback through E-Cadherin Promotes Direction Sensing during Collective Cell Migration , 2014, Cell.

[52]  Taekjip Ha,et al.  Defining Single Molecular Forces Required to Activate Integrin and Notch Signaling , 2013, Science.

[53]  Michael Krieg,et al.  Mechanical Control of the Sense of Touch by β Spectrin , 2014, Nature Cell Biology.

[54]  Jessica R. Harrell,et al.  Apical constriction: a cell shape change that can drive morphogenesis. , 2010, Developmental biology.

[55]  M. Rief,et al.  Extracellular rigidity sensing by talin isoform–specific mechanical linkages , 2015, Nature Cell Biology.

[56]  Christopher S. Chen,et al.  Cells lying on a bed of microneedles: An approach to isolate mechanical force , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[57]  Matthew E. Berginski,et al.  Construction, imaging, and analysis of FRET-based tension sensors in living cells. , 2015, Methods in cell biology.

[58]  M. Rief,et al.  The Complex Folding Network of Single Calmodulin Molecules , 2011, Science.

[59]  K. Salaita,et al.  Integrin-generated forces lead to streptavidin-biotin unbinding in cellular adhesions. , 2014, Biophysical journal.

[60]  M. Sheetz,et al.  Force of single kinesin molecules measured with optical tweezers. , 1993, Science.

[61]  Carsten Grashoff,et al.  Generation and analysis of biosensors to measure mechanical forces within cells. , 2013, Methods in molecular biology.

[62]  S. Sen,et al.  Matrix Elasticity Directs Stem Cell Lineage Specification , 2006, Cell.