Signal Transduction Through Trimeric G Proteins in Megakaryoblastic Cell Lines

The biogenesis of trimeric G proteins was investigated by measurement of the expression of alpha-subunits in the megakaryoblastic cell lines MEG-01, DAMI, and CHRF-288-11, representing stages of increasing maturation, and compared with platelets. Megakaryoblasts and platelets contained approximately equal amounts of Gi alpha-1/2, G (i) alpha-3, Gq alpha, and G12 alpha protein. Maturation was accompanied by (1) downregulation of mRNA for Gs alpha and disappearance of iloprost-induced Ca mobilization, (2) upregulation of the long form of Gs alpha protein (Gs alpha-L) and an increase in iloprost-induced cAMP formation, and (3) upregulation of G16 alpha mRNA and G16 alpha protein and appearance of thromboxane A (2) -induced signaling (Ca mobilization and stimulation of prostaglandin I2 -induced cAMP formation). G2 alpha protein was absent in the megakaryoblasts despite weak expression of G2 alpha mRNA in DAMI and relatively high levels of G2 alpha mRNA and G2 alpha protein in platelets. These findings reveal major changes in G protein-mediated signal transduction during megakaryocytopoiesis and indicate that G16 alpha couples the thromboxane receptor to phospholipase Cbeta. (Arterioscler Thromb Vasc Biol. 1997;17:1830-1836.)