The small GTPase Rap1b regulates the cross talk between platelet integrin alpha2beta1 and integrin alphaIIbbeta3.
The involvement of the small GTPase Rap1b in platelet integrin alpha2beta1-dependent outside-in signaling was investigated. Platelet adhesion to 4 different specific ligands for integrin alpha2beta1, monomeric collagen, decorin, and collagen-derived peptides CB8(II) and CB11(II), induced a robust and rapid activation of Rap1b. This process did not require secreted ADP or thromboxane A2 production but was critically regulated by phospholipase C (PLC)-derived second messengers. Both Ca2+ and protein kinase C were found to organize independent but additive pathways for Rap1b activation downstream of integrin-alpha2beta1, which were completely blocked by inhibition of PLC with U73122. Moreover, integrin alpha2beta1 engagement failed to trigger Rap1b activation in murine platelets lacking CalDAG-GEFI, a guanine nucleotide exchange factor regulated by Ca2+ and diacylglycerol, despite normal phosphorylation and activation of PLCgamma2. In addition, CalDAG-GEFI-deficient platelets showed defective integrin alpha2beta1-dependent adhesion and spreading. We found that outside-in signaling through integrin alpha2beta1 triggered inside-out activation of integrin alphaIIbbeta3 and promoted fibrinogen binding. Similarly to Rap1b stimulation, this process occurred downstream of PLC activation and was dramatically impaired in murine platelets lacking the Rap1 exchange factor CalDAG-GEFI. These results demonstrate that Rap1b is an important element in integrin-dependent outside-in signaling during platelet adhesion and regulates the cross talk between adhesive receptors.
Bernardi
Bruno
B
Department of Biochemistry, University of Pavia, via Bassi 21, 27100 Pavia, Italy.
Guidetti
Gianni F
GF
Campus
Francesca
F
Crittenden
Jill R
JR
Graybiel
Ann M
AM
Balduini
Cesare
C
Torti
Mauro
M
D004789:Enzyme Activation; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D038982:Integrin alpha2beta1 / Q000097:blood / Q000502:physiology*; D007700:Kinetics; D010973:Platelet Adhesiveness / Q000502:physiology*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000502:physiology*; D020239:Receptor Cross-Talk / Q000502:physiology*; D015290:Second Messenger Systems / Q000502:physiology; D010738:Type C Phospholipases / Q000097:blood; D020668:rap GTP-Binding Proteins / Q000097:blood / Q000378:metabolism*
D038982:Integrin alpha2beta1; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D005340:Fibrinogen; D010738:Type C Phospholipases; C116753:RAP1B protein, human; D020668:rap GTP-Binding Proteins