Abstract
Integrin-based adhesion to the extracellular matrix (ECM) plays critical roles in controlling differentiation, survival, and motility of epithelial cells. Cells attach to the ECM via dynamic structures called focal adhesions (FA). FA undergo constant remodeling mediated by vesicle trafficking and fusion. A soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein α (αSNAP) is an essential mediator of membrane fusion; however, its roles in regulating ECM adhesion and cell motility remain unexplored. In this study, we found that siRNA-mediated knockdown of αSNAP induced detachment of intestinal epithelial cells, whereas overexpression of αSNAP increased ECM adhesion and inhibited cell invasion. Loss of αSNAP impaired Golgi-dependent glycosylation and trafficking of β1 integrin and decreased phosphorylation of focal adhesion kinase (FAK) and paxillin resulting in FA disassembly. These effects of αSNAP depletion on ECM adhesion were independent of apoptosis and NSF. In agreement with our previous reports that Golgi fragmentation mediates cellular effects of αSNAP knockdown, we found that either pharmacologic or genetic disruption of the Golgi recapitulated all the effects of αSNAP depletion on ECM adhesion. Furthermore, our data implicates β1 integrin, FAK, and paxillin in mediating the observed pro-adhesive effects of αSNAP. These results reveal novel roles for αSNAP in regulating ECM adhesion and motility of epithelial cells.
Highlights
Vesicle trafficking plays important roles in regulating cell adhesion and motility
extracellular matrix (ECM) adhesion and motility of epithelial cells are regulated by elaborate vesicle trafficking/fusion machinery that is critical for assembly and remodeling of matrix adhesion complexes
The present study identifies novel functions of a key membrane fusion protein, ␣soluble NSF-attachment proteins (SNAPs), in controlling ECM adhesion and invasion of human intestinal epithelial cells
Summary
Results: Depletion of the membrane fusion protein, N-ethylmaleimide-sensitive factor attachment protein ␣ (␣SNAP), induced cell detachment, whereas overexpression of this vesicle regulator increased cell-matrix adhesion. A soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein ␣ (␣SNAP) is an essential mediator of membrane fusion; its roles in regulating ECM adhesion and cell motility remain unexplored. Our data implicates 1 integrin, FAK, and paxillin in mediating the observed pro-adhesive effects of ␣SNAP These results reveal novel roles for ␣SNAP in regulating ECM adhesion and motility of epithelial cells. We identified ␣SNAP as a critical positive regulator of ECM adhesion of human epithelial cells via multiple mechanisms that control FA assembly and Golgi-dependent maturation and trafficking of 1 integrin
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