Coupling of vinculin to F-actin demands Syndecan-4 proteoglycan

- Syndecan-4 knockdown in endothelial cells leads to decoupling of vinculin from F-actin filaments.
- Syndecan-4 knockdown increases filopodia-like structures in the cells.
- Syndecan-4 knockdown alters β1 integrin and fibronectin expression and localization.
- Syndecan-4 loss impairs cell-ECM interaction leading to decrease in cell adhesion and increased cell mobility.
- Syndecan-4 is as a central mediator in focal adhesion complex bridging fibronectin, integrin and intracellular components.

Syndecans are heparan sulfate proteoglycans characterized as transmembrane receptors that act cooperatively with the cell surface and extracellular matrix proteins. Syn4 knockdown was performed in order to address its role in endothelial cells (EC) behavior. Normal EC and shRNA-Syn4-EC cells were studied comparatively using complementary confocal, super-resolution and non-linear microscopic techniques. Confocal and super-resolution microscopy revealed that Syn4 knockdown alters the level and arrangement of essential proteins for focal adhesion, evidenced by the decoupling of vinculin from F-actin filaments. Furthermore, Syn4 knockdown alters the actin network leading to filopodial protrusions connected by VE-cadherin-rich junction. shRNA-Syn4-EC showed reduced adhesion and increased migration. Also, Syn4 silencing alters cell cycle as well as cell proliferation. Moreover, the ability of EC to form tube-like structures in matrigel is reduced when Syn4 is silenced. Together, the results suggest a mechanism in which Syndecan-4 acts as a central mediator that bridges fibronectin, integrin and intracellular components (actin and vinculin) and once silenced, the cytoskeleton protein network is disrupted. Ultimately, the results highlight Syn4 relevance for balanced cell behavior.