The fibronectin-binding integrins α5β1 and αvβ3 differentially modulate RhoA-GTP loading, organization of cell matrix adhesions, and fibronectin fibrillogenesis

Abstract

We have studied the formation of different types of cell matrix adhesions in cells that bind to fibronectin via either α5β1 or αvβ3. In both cases, cell adhesion to fibronectin leads to a rapid decrease in RhoA activity. However, α5β1 but not αvβ3 supports high levels of RhoA activity at later stages of cell spreading, which are associated with a translocation of focal contacts to peripheral cell protrusions, recruitment of tensin into fibrillar adhesions, and fibronectin fibrillogenesis. Expression of an activated mutant of RhoA stimulates αvβ3-mediated fibrillogenesis. Despite the fact that α5β1-mediated adhesion to the central cell-binding domain of fibronectin supports activation of RhoA, other regions of fibronectin are required for the development of α5β1-mediated but not αvβ3-mediated focal contacts. Using chimeras of β1 and β3 subunits, we find that the extracellular domain of β1 controls RhoA activity. By expressing both β1 and β3 at high levels, we show that β1-mediated control of the levels of β3 is important for the distribution of focal contacts. Our findings demonstrate that the pattern of fibronectin receptors expressed on a cell dictates the ability of fibronectin to stimulate RhoA-mediated organization of cell matrix adhesions.