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, Gi alpha-3, Gq alpha, and G12 alpha protein. Maturation was accompanied by (1) downregulation of mRNA for Gs alpha and disappearance of iloprost-induced Ca2+ 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 A2-induced signaling (Ca2+ mobilization and stimulation of prostaglandin I2-induced cAMP formation). Gz alpha protein was absent in the megakaryoblasts despite weak expression of Gz alpha mRNA in DAMI and relatively high levels of Gz alpha mRNA and Gz 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 C beta.
van der Vuurst
H
H
Department of Hematology, University Hospital Utrecht, Netherlands.
van Willigen
G
G
van Spronsen
A
A
Hendriks
M
M
Donath
J
J
Akkerman
J W
JW
D001692:Biological Transport / Q000187:drug effects; D001792:Blood Platelets / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D016922:Cellular Senescence / Q000502:physiology; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D000242:Cyclic AMP / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D016285:Iloprost / Q000494:pharmacology; D008533:Megakaryocytes / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D013928:Thromboxane A2 / Q000494:pharmacology
D012333:RNA, Messenger; D013928:Thromboxane A2; D000242:Cyclic AMP; D019204:GTP-Binding Proteins; D016285:Iloprost; D002118:Calcium