Orai1 is required for Ca²⁺-dependent plasma membrane repair and mechanoadaptation

Ca²⁺-dependent repair of plasma membrane breaches is essential for animal cell viability. An initial passive influx of extracellular Ca²⁺ triggers the formation of a protein plug that rapidly seals breaches. However, the mechanism of extracellular Ca²⁺ requirement for subsequent repair remains undefined. EHD2 protein stabilizes the plasma membrane caveolae, which sustain membrane repair, and maintains high surface levels of the caveolae-resident Ca²⁺ channel Orai1. We establish the requirement of both Orai1 and EHD2 for repair of plasma membrane lesions induced by mechanical injury or by a model bacterial pore-forming toxin. We demonstrate rapid EHD2 recruitment and Orai1-mediated Ca²⁺ entry at plasma membrane sites of localized mechanical stimulus, the latter requiring EHD2 and CAV1. EHD2 and Orai1 are necessary for mechanosensitive YAP/TAZ-TEAD activation and positive feedback for CAV1 expression that promotes membrane repair. Our studies establish EHD2 and Orai1 as novel components of mammalian plasma membrane repair and mechanoadaptation.