The role of actin-binding protein 280 in integrin-dependent mechanoprotection

To survive in a mechanically active environment, cells must adapt to variations of applied membrane tension, A collagen-coated magnetic bead model was used to apply forces directly to the actin cytoskeleton through integrin receptors, We demonstrate here that by a calcium-dependent mechanism, human fibroblasts reinforce locally their connection with extracellular adhesion sites by inducing actin assembly and by recruiting actin-binding protein 280 (ABP-280) into cortical adhesion complexes, ABP-280 was phosphorylated on serine residues as a result of force application, This phosphorylation and the force-induced actin reorganization were largely abrogated by inhibitors of protein kinase C. In a human melanoma cell line that does not express ABP-280, actin accumulation could not be induced by force, whereas in stable transfectants expressing ABP-280, force-induced actin accumulation was similar to human fibroblasts, Cortical actin assembly played a role in regulating the activity of stretch-activated, calcium permeable channels (SAG) since sustained force application desensitized SAC to subsequent force applications, and the decrease in stretch sensitivity was reversed after treatment with cytochalasin D. ABP-280-deficient cells showed a >90% increase in cell death compared with ABP-280+ve cells after force application. We conclude that ABP-280 plays an important role in mechanoprotection by reinforcing the membrane cortex and desensitizing SACs.