title: Optogenetic control of cellular forces and mechanotransduction.
abstract: Contractile forces are the end effectors of cell migration, division, morphogenesis, wound healing and cancer invasion. Here we report optogenetic tools to upregulate and downregulate such forces with high spatiotemporal accuracy. The technology relies on controlling the subcellular activation of RhoA using the CRY2/CIBN light-gated dimerizer system. We fused the catalytic domain (DHPH domain) of the RhoA activator ARHGEF11 to CRY2-mCherry (optoGEF-RhoA) and engineered its binding partner CIBN to bind either to the plasma membrane or to the mitochondrial membrane. Translocation of optoGEF-RhoA to the plasma membrane causes a rapid and local increase in cellular traction, intercellular tension and tissue compaction. By contrast, translocation of optoGEF-RhoA to mitochondria results in opposite changes in these physical properties. Cellular changes in contractility are paralleled by modifications in the nuclear localization of the transcriptional regulator YAP, thus showing the ability of our approach to control mechanotransductory signalling pathways in time and space.
authors.affiliation: Institute for Bioengineering of Catalonia, Barcelona 08028, Spain.
authors.forename: Léo
authors.initials: L
authors.lastname: Valon
authors.affiliation: Institute for Bioengineering of Catalonia, Barcelona 08028, Spain.
authors.forename: Ariadna
authors.initials: A
authors.lastname: Marín-Llauradó
authors.affiliation: MRC Laboratory for Molecular Cell Biology, University College London, Gower Street, London WC1E 6BT, UK.|London Centre for Nanotechnology, London WC1H 0AH, UK.
authors.forename: Thomas
authors.initials: T
authors.lastname: Wyatt
authors.affiliation: London Centre for Nanotechnology, London WC1H 0AH, UK.|Department of Cell and Developmental Biology and Institute for the Physics of Living Systems, University College London, London WC1E 6BT, UK.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Charras
authors.affiliation: Institute for Bioengineering of Catalonia, Barcelona 08028, Spain.|Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain.|Institució Catalana de Recerca i Estudis Avançats (ICREA), 08010 Barcelona, Spain.|Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina, Barcelona 08028, Spain.
authors.forename: Xavier
authors.initials: X
authors.lastname: Trepat
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement / Q000502:physiology*; D056931:Cryptochromes / Q000235:genetics / Q000378:metabolism; D004285:Dogs; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D061985:Madin Darby Canine Kidney Cells; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D051336:Mitochondrial Membranes / Q000378:metabolism; D062308:Optogenetics / Q000379:methods*; D011485:Protein Binding; D021381:Protein Transport; D064067:Rho Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D056931:Cryptochromes; D008164:Luminescent Proteins; D064067:Rho Guanine Nucleotide Exchange Factors; D020742:rhoA GTP-Binding Protein; D000097573:Red Fluorescent Protein