Hippo effectors YAP/TAZ act as on-off mechanosensing switches by sensing modifications in extracellular matrix (ECM) composition and mechanics. The regulation of their activity has been described by a hierarchical model in which elements of Hippo pathway are under the control of focal adhesions (FAs). Here we unveil the molecular mechanism by which cell spreading and RhoA GTPase activity control FA formation through YAP to stabilize the anchorage of the actin cytoskeleton to the cell membrane. This mechanism requires YAP co-transcriptional function and involves the activation of genes encoding for integrins and FA docking proteins. Tuning YAP transcriptional activity leads to the modification of cell mechanics, force development and adhesion strength, and determines cell shape, migration and differentiation. These results provide new insights into the mechanism of YAP mechanosensing activity and qualify this Hippo effector as the key determinant of cell mechanics in response to ECM cues.

CEITEC MU Masaryk University CZ 65691 Brno Czech Republic

Department of Biomaterials Science Institute of Dentistry University of Turku FI 20014 Turku Finland

Faculty of Medicine and Dentistry Institute of Molecular and Translational Medicine Palacky University Olomouc Hněvotínská 1333 5 CZ 77515 Olomouc Czech Republic

Faculty of Medicine Department of Biology Masaryk University CZ 62500 Brno Czech Republic

Faculty of Pharmacy Division of Pharmaceutical Biosciences University of Helsinki Viikinkaari 5 E FI 00014 Helsinki Finland

Institute of Advanced Biomedical Engineering and Science Tokyo Women's Medical University 8 1 Kawada Cho Shinjuku Ku Tokyo JP 162 8666 Japan

International Clinical Research Center St Anne's University Hospital CZ 65691 Brno Czech Republic

Ket Lab Italian Space Agency Via del Politecnico snc 00133 Rome Italy

Laboratory of Bio inspired and Graphene Nanomechanics Department of Civil Environmental and Mechanical Engineering University of Trento 1 38123 Trento Italy

School of Engineering and Materials Science Queen Mary University of London Mile End Road UK E1 4NS London UK

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