Integrin activation.
Agonist stimulation of integrin receptors, composed of transmembrane alpha and beta subunits, leads cells to regulate integrin affinity ('activation'), a process that controls cell adhesion and migration, and extracellular matrix assembly. A final step in integrin activation is the binding of talin to integrin beta cytoplasmic domains. We used forward, reverse and synthetic genetics to engineer and order integrin activation pathways of a prototypic integrin, platelet alphaIIbbeta3. PMA activated alphaIIbbeta3 only after expression of both PKCalpha (protein kinase Calpha) and talin at levels approximating those in platelets. Inhibition of Rap1 GTPase reduced alphaIIbbeta3 activation, whereas expression of constitutively active Rap1A(G12V) bypassed the requirement for PKCalpha. Overexpression of a Rap effector, RIAM (Rap1-GTP-interacting adaptor molecule), activated alphaIIbbeta3 and bypassed the requirement for PKCalpha and Rap1. In addition, shRNA (short hairpin RNA)-mediated knockdown of RIAM blocked talin interaction with and activation of integrin alphaIIbbeta3. Rap1 activation caused the formation of an 'activation complex' containing talin and RIAM that redistributed to the plasma membrane and activated alphaIIbbeta3. The central finding was that this Rap1-induced formation of an 'integrin activation complex' leads to the unmasking of the integrin-binding site on talin, resulting in integrin activation.
Banno
Asoka
A
Department of Medicine, University of California, San Diego, 9500 Gilman Drive, MC 0726, San Diego, CA 92093-0726, USA.
Ginsberg
Mark H
MH
D048868:Adaptor Proteins, Signal Transducing / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000819:agonists / Q000378:metabolism*; D051571:Protein Kinase C-alpha / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D016608:Talin / Q000378:metabolism; D020827:rap1 GTP-Binding Proteins / Q000378:metabolism
C493340:APBB1IP protein, human; D048868:Adaptor Proteins, Signal Transducing; D008565:Membrane Proteins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D016608:Talin; D051571:Protein Kinase C-alpha; D020827:rap1 GTP-Binding Proteins