Regulation of β1 Integrin recycling, cell migration and focal adhesion turnover by CD13

Abstract CD13 is a multifunctional cell surface peptidase that is expressed on a variety of cells where we have shown that it modulates receptor-mediated endocytosis and ligand internalization to control downstream signaling pathways. In fibroblasts and epithelial cells, CD13 is localized with β1-integrin at focal adhesions (FAs), the sites of communication between the extracellular matrix and the actin cytoskeleton, prompting our exploration of potential contributions of CD13 focal adhesion turnover, integrin endocytosis and trafficking. Phenotypically, FAs in CD13KO fibroblasts are elongated and irregular with displaced FA accessory proteins, markedly reduced actin stress fibers and fewer microtubule extensions, consistent with a link between CD13 and the control of FA dynamics. In wild type cells, CD13 and β1-integrin co-internalize, traffic to EEA1+ Rab5+ early endosomes and recycle to the plasma membrane via Rab11a+ recycling endosomes. Pulse-chase assays with wild type and CD13 phospho tyrosine mutant confirmed that tyrosine phosphorylated CD13 must associate with the ARF6 GTPase for β1-integrin recycling to the membrane to occur. Conversely, the absence of CD13 results in trafficking of internalized β1-integrin from early endosomes to Rab7+ late endosomes and lysosomes and a failure to return to the cell surface. Functionally, in migrating cells CD13 accumulates at the leading edge and co-localizes with IQGAP1 and focal adhesion protein, Paxillin to regulate cell migration and adhesion. In conclusion, CD13 is responsible for proper localization of focal adhesion proteins, coordination of integrin recycling and focal adhesion dynamics, thereby regulating integrin signal transduction and cell motility.