Integrin-dependent force transmission to the extracellular matrix by α-actinin triggers adhesion maturation

Focal adhesions are mechanosensitive elements that enable mechanical communication between cells and the extracellular matrix. Here, we demonstrate a major mechanosensitive pathway in which α-actinin triggers adhesion maturation by linking integrins to actin in nascent adhesions. We show that depletion of the focal adhesion protein α-actinin enhances force generation in initial adhesions on fibronectin, but impairs mechanotransduction in a subsequent step, preventing adhesion maturation. Expression of an α-actinin fragment containing the integrin binding domain, however, dramatically reduces force generation in depleted cells. This behavior can be explained by a competition between talin (which mediates initial adhesion and force generation) and α-actinin for integrin binding. Indeed, we show in an in vitro assay that talin and α-actinin compete for binding to β₃ integrins, but cooperate in binding to β₁ integrins. Consistently, we find opposite effects of α-actinin depletion and expression of mutants on substrates that bind β₃ integrins (fibronectin and vitronectin) versus substrates that only bind β₁ integrins (collagen). We thus suggest that nascent adhesions composed of β₃ integrins are initially linked to the actin cytoskeleton by talin, and then α-actinin competes with talin to bind β₃ integrins. Force transmitted through α-actinin then triggers adhesion maturation. Once adhesions have matured, α-actinin recruitment correlates with force generation, suggesting that α-actinin is the main link transmitting force between integrins and the cytoskeleton in mature adhesions. Such a multistep process enables cells to adjust forces on matrices, unveiling a role of α-actinin that is different from its well-studied function as an actin cross-linker.