Disruption of β2-integrin-cytoskeleton coupling abolishes the signaling capacity of these integrins on granulocytes

Integrin-dependent adhesion and dynamic modulations of the actin network are prerequisites for normal cell locomotion. To investigate whether the actin microfilamentous system does play a role in regulation of beta(2)-integrin-induced signalling, we pretreated granulocytes with staurosporine, a well-known protein kinase inhibitor that has also been shown to disrupt the cytoskeleton of intact cells. Pretreatment with staurosporine completely inhibited the beta(2-)integrin-induced Ca2+ signal and also its ability to trigger actin polymerisation. This inhibition was not related to phosphorylation of the CD18-chain of the beta(2)-integrin, nor to inhibition of protein kinases. Instead, association of beta(2)-integrins with the cortical cytoskeleton, which was observed in untreated cells, was abolished after exposure to staurosporine, indicating that beta(2)-integrin signalling depends on integrin-cytoskeleton interaction. These results suggest not only that the actin network provides an adhesive link to the extracellular matrix and a driving force for the locomotory response, but also that it participates in regulation of beta(2)-integrin signalling during granulocyte locomotion. (C) 1999 Academic Press.