degradation of internalized αvβ5 integrin is controlled by upar bound upa effect on β1 integrin activity and α-sma stress fiber assembly退化内化αvβ5整合素是由upar绑定upa影响β1整合素活动和α-sma应力纤维组装.pdfVIP

Degradation of Internalized avb5 Integrin Is Controlled by uPAR Bound uPA: Effect on b1 Integrin Activity and a- SMA Stress Fiber Assembly 1 1 1 1 2 Lingyan Wang , Benjamin S. Pedroja , Erin E. Meyers , Angelo L. Garcia , Sally S. Twining , Audrey M. Bernstein1* 1 Department of Ophthalmology, Mount Sinai School of Medicine, New York, New York, United States of America, 2 Department of Biochemistry, Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America Abstract Myofibroblasts (Mfs) that persist in a healing wound promote extracellular matrix (ECM) accumulation and excessive tissue contraction. Increased levels of integrin avb5 promote the Mf phenotype and other fibrotic markers. Previously we reported that maintaining uPA (urokinase plasminogen activator) bound to its cell-surface receptor, uPAR prevented TGFb-induced Mf differentiation. We now demonstrate that uPA/uPAR controls integrin b5 protein levels and in turn, the Mf phenotype. When cell-surface uPA was increased, integrin b5 levels were reduced (61%). In contrast, when uPA/uPAR was silenced, integrin b5 total and cell-surface levels were increased (2–4 fold). Integrin b5 accumulation resulted from a significant decrease in b5 ubiquitination leading to a decrease in the degradation rate of internalized b5. uPA-silencing also induced a- SMA stress fiber organization in cells that were seeded on collagen, increased cell area (1.7 fold), and increased integrin b1 binding to the collagen matrix, with reduced activation of b1. Elevated cell-surface integrin b5 was necessary for these changes after uPA-silencing since blocking avb5 function reversed these effects. Our data support a novel mechanism by which downregulation of