Activation of protein kinase C in rat basophilic leukemia cells stimulates increased production of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate: correlation with actin polymerization.
Cross-linking of the immunoglobulin E receptor on rat basophilic leukemia (RBL)1 cells by multivalent antigen activates phosphatidylinositol (PI) kinase and phosphatidylinositol 4-phosphate (PIP) kinase leading to the increased production of PIP and phosphatidylinositol 4,5-bisphosphate (PIP2). Activators of protein kinase C (PKC), such as phorbol myristate acetate (PMA) and the synthetic diacylglycerol, 1,2-dioctanoyl-sn-glycerol (diC8), were found to have the same effect even though PMA and diC8 do not cause the activation of phospholipase C. Although the kinetics are different depending on the stimulant, activation of PKC using multivalent antigen, PMA or diC8 also causes the polymerization of actin and an increase in the F-actin content of the cells. In all cases, a good correlation was observed between F-actin levels, activation of PI and PIP kinases, and the increased production of PIP and PIP2. However, in the case of antigen, there is no correlation between actin polymerization and the total amount of PIP and PIP2. Staurosporine, an inhibitor of protein kinases, blocks the F-actin response and the increased synthesis of PIP and PIP2 with similar dose dependencies. Furthermore, depletion of PKC activity through long-term exposure to PMA, inhibited both the F-actin response and the increased synthesis of PIP and PIP2 induced by either DNP-BSA or diC8. These results suggest that activation of PKC precedes the activation of PI and PIP kinases and that under certain circumstances activation of the kinases and the increased synthesis of PIP and PIP2 may be involved in the polymerization of actin in RBL cells, possibly through the interaction of the polyphosphoinositides with actin-binding proteins such as gelsolin and profilin.
Apgar
J R
JR
Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, California 92037, USA.
D000199:Actins / Q000378:metabolism*; D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D001704:Biopolymers; D004075:Diglycerides / Q000494:pharmacology; D004140:Dinitrophenols / Q000276:immunology; D004789:Enzyme Activation; D007152:Immunologic Capping; D015471:Leukemia, Basophilic, Acute / Q000201:enzymology*; D009363:Neoplasm Proteins / Q000502:physiology*; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D051381:Rats; D017455:Receptors, IgE / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000276:immunology; D015398:Signal Transduction; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000199:Actins; D000470:Alkaloids; D001704:Biopolymers; D004075:Diglycerides; D004140:Dinitrophenols; D009363:Neoplasm Proteins; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; D017455:Receptors, IgE; C034638:dinitrophenyl-bovine serum albumin; C037178:phosphatidylinositol 4-phosphate; C044295:1,2-dioctanoylglycerol; D012710:Serum Albumin, Bovine; D011493:Protein Kinase C; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate