RGS/Gi2alpha interactions modulate platelet accumulation and thrombus formation at sites of vascular injury.
Although much is known about extrinsic regulators of platelet function such as nitric oxide and prostaglandin I(2) (PGI(2)), considerably less is known about intrinsic mechanisms that prevent overly robust platelet activation after vascular injury. Here we provide the first evidence that regulators of G-protein signaling (RGS) proteins serve this role in platelets, using mice with a G184S substitution in G(i2α) that blocks RGS/G(i2) interactions to examine the consequences of lifting constraints on G(i2)-dependent signaling without altering receptor:effector coupling. The results show that the G(i2α)(G184S) allele enhances platelet aggregation in vitro and increases platelet accumulation after vascular injury when expressed either as a global knock-in or limited to hematopoietic cells. Biochemical studies show that these changes occur in concert with an attenuated rise in cyclic adenosine monophosphate levels in response to prostacyclin and a substantial increase in basal Akt activation. In contrast, basal cyclic adenosine monophosphate (cAMP) levels, agonist-stimulated increases in [Ca(++)](i), Rap1 activation, and α-granule secretion were unaffected. Collectively, these observations (1) demonstrate an active role for RGS proteins in regulating platelet responsiveness, (2) show that this occurs in a pathway-selective manner, and (3) suggest that RGS proteins help to prevent unwarranted platelet activation as well as limiting the magnitude of the normal hemostatic response.
Signarvic
Rachel S
RS
Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Cierniewska
Aleksandra
A
Stalker
Timothy J
TJ
Fong
Karen P
KP
Chatterjee
Manash S
MS
Hess
Paul R
PR
Ma
Peisong
P
Diamond
Scott L
SL
Neubig
Richard R
RR
Brass
Lawrence F
LF
D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D000242:Cyclic AMP / Q000494:pharmacology; D005260:Female; D051960:GTP-Binding Protein alpha Subunit, Gi2 / Q000235:genetics / Q000378:metabolism*; D015151:Immunoblotting; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation / Q000235:genetics; D010766:Phosphorylation; D015539:Platelet Activation; D010974:Platelet Aggregation; D010976:Platelet Count; D051057:Proto-Oncogene Proteins c-akt; D020710:RGS Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013927:Thrombosis / Q000378:metabolism*; D057772:Vascular System Injuries / Q000378:metabolism / Q000473:pathology*
D020710:RGS Proteins; D000242:Cyclic AMP; D051057:Proto-Oncogene Proteins c-akt; D051960:GTP-Binding Protein alpha Subunit, Gi2; D002118:Calcium