AbstractThe mechanically activated Piezo channel plays a versatile role in conferring mechanosensitivity to various cell types. However, how it incorporates its intrinsic mechanosensitivity and cellular components to effectively sense long-range mechanical perturbation across a cell remains elusive. Here we show that Piezo1 is biochemically and functionally tethered to the actin cytoskeleton via the E-cadherin-β-catenin mechanotransduction complex, whose perturbation significantly impairs Piezo1-mediated responses. Mechanistically, the adhesive extracellular domain of E-cadherin interacts with the cap domain of Piezo1 that controls the transmembrane gate, while its cytosolic tail might interact with the cytosolic domains of Piezo1 that are in close proximity to its intracellular gates, allowing a direct focus of adhesion-cytoskeleton-transmitted force for gating. Specific disruption of the intermolecular interactions prevents cytoskeleton-dependent gating of Piezo1. Thus, we propose a force-from-filament model to complement the previously suggested force-from-lipids model for mechanogating of Piezo channels, enabling them to serve as versatile and tunable mechanotransducers.HighlightsRevealed biochemical and functional interactions between Piezo1 and the E-cadherin-β-catenin-F-actin mechanotransduction complex.Identified critical mechanogating domains of Piezo1 as E-cadherin binding domains.Specific disruption of the intermolecular interactions between Piezo1 and E-cadherin prevents cytoskeleton-dependent gating of Piezo1.Proposed a tether model for mechanogating of Piezo channels.