Regulation of urokinase receptor function and pericellular proteolysis by the integrin α₅β₁

 

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Summary

Interactions between the uPA receptor (uPAR) and various inte-grins, including α₅β₁, are known to modulate integrin-dependent cell adhesion, and we have shown that the integrin-associated tetraspanin protein CD82 down-regulates uPAR-dependent plasminogen activation by affecting α₅β₁ cellular localisation. Here we have investigated whether overexpression of α₅β₁ directly affects uPAR-dependent pericellular proteolysis. CHO cells overexpressing α₅β₁ were found to activate plasminogen at a rate up to 18-fold faster than B2CHO cells which are α₅-deficient. This effect was dependent on the activation state of α₅β₁, as it was maximal in the presence of Mn²⁺. To determine the role of uPAR-α₅β₁ interactions in this effect, we determined the adhesion of these cells to immobilised soluble uPAR (suPAR). Neither cell-type was found to adhere to suPAR, but both cell types were found to adhere to an anti-uPAR monoclonal antibody in a uPAR- and integrin-dependent manner. This adhesion was 10-fold greater in the absence of α₅β₁, possibly implicating the involvement of non-α₅-integrins. Soluble forms of the various components were used to investigate the molecular basis of these effects, but no direct interactions could be demonstrated between α₅β₁ and either uPAR, uPA or uPA-uPAR complex. This suggests that assembly of these components on the plasma membrane is required to influence uPAR function, increasing uPAR-dependent pericellular proteolysis and decreasing uPAR-dependent cell adhesion. These interactions may be modified by other integrins, suggesting a complex interplay between uPAR and integrins on the cell surface with the potential to regulate invasive cell migration.

• References

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