EphA2 signaling within integrin adhesions regulates fibrillar adhesion elongation and fibronectin deposition

The multifunctional glycoprotein fibronectin influences several crucial cellular processes and contributes to multiple pathologies. While a link exists between fibronectin-associated pathologies and the receptor tyrosine kinase EphA2, the mechanism by which EphA2 promotes fibronectin matrix remodeling remains unknown. We previously demonstrated that EphA2 deletion reduces smooth muscle fibronectin deposition and blunts fibronectin deposition in atherosclerosis without influencing fibronectin expression. We now show that EphA2 expression is required for contractility-dependent elongation of tensin- and α5β1 integrin-rich fibrillar adhesions that drive fibronectin fibrillogenesis. Mechanistically, EphA2 localizes to integrin adhesions where focal adhesion kinase mediates ligand-independent Y772 phosphorylation, and mutation of this site significantly blunts fibrillar adhesion length. EphA2 deficiency decreases smooth muscle cell contractility by enhancing p190RhoGAP activation and reducing RhoA activity, whereas stimulating RhoA signaling in EphA2 deficient cells rescues fibrillar adhesion elongation. Together, these data identify EphA2 as a novel regulator of fibrillar adhesion elongation and provide the first data identifying a role for EphA2 signaling in integrin adhesions. [Display omitted] •EphA2 expression is required for efficient elongation of tensin- and α5β1-rich fibrillar adhesions in vascular smooth muscle cells.•EphA2 undergoes ligand-independent Y772 phosphorylation within integrin adhesions through focal adhesion kinase-dependent transphosphorylation.•EphA2 Y772 phosphorylation, but not EphA2 ligand-binding, promotes fibrillar adhesion elongation and fibronectin deposition by enhancing RhoA-mediated contractility.•EphA2 expression limits p190RhoGAP activation to promote contractility-dependent fibrillar adhesion formation and fibronectin deposition.