Platelet Gs hypofunction and abnormal morphology resulting from a heterozygous RGS2 mutation.
BACKGROUND
Regulator of G-protein signaling (RGS) 2 negatively regulates Gs signaling by inhibiting the activation of adenylyl cyclase (AC). RGS2 mRNA contains four translation initiation sites, leading to four isoforms with different abilities to inhibit AC activity; the largest isoform is the most pronounced inhibitor. A role for RGS2 in platelets is not known.


OBJECTIVE
To describe a heterozygous RGS2 mutation (G23D) in three related patients, leading to Gs hypofunction in their platelets, and to study the mechanism behind the effect of the RGS2 mutation on platelet function and morphology.


METHODS
Gs signaling was studied ex vivo in platelets and in vitro in transfected cells. Translation initiation was evaluated in vitro, and the interaction of wild-type and G23D RGS2 with AC was unraveled via immunoprecipitation. Platelet granule content was analyzed with proteomics.


RESULTS
The mutation leads to reduced cAMP production after stimulation of Gs-coupled receptors. The largest RGS2 isoforms, with strong AC inhibitor activity, are enriched when the mutation is present, as compared with wild-type RGS2. Moreover, the mutation results in a stronger interaction of RGS2 with AC. G23D RGS2 carriers have enlarged, round platelets with abnormal alpha-granules. Proteomics of the platelet releasate revealed altered expression of some proteins involved in actin assembly, and carriers seemed to have a reduced platelet shape change.


CONCLUSIONS
We present the first platelet Gs signaling defect caused by a heterozygous RGS2 variant that results in a unique mutational mechanism, such as the differential use of translation initiation sites resulting in different functional RGS2 isoforms.
Noé
L
L
Centre for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium.
Di Michele
M
M
Giets
E
E
Thys
C
C
Wittevrongel
C
C
De Vos
R
R
Overbergh
L
L
Waelkens
E
E
Jaeken
J
J
Van Geet
C
C
Freson
K
K
D000067956:Adenylyl Cyclase Inhibitors; D001792:Blood Platelets / Q000378:metabolism* / Q000473:pathology / Q000502:physiology; D048430:Cell Shape / Q000235:genetics; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006579:Heterozygote; D006801:Humans; D020125:Mutation, Missense; D014176:Protein Biosynthesis; D020033:Protein Isoforms; D020710:RGS Proteins / Q000235:genetics / Q000502:physiology*; D015398:Signal Transduction
D000067956:Adenylyl Cyclase Inhibitors; D020033:Protein Isoforms; D020710:RGS Proteins; C087182:RGS2 protein, human; D019204:GTP-Binding Proteins