Point mutation in the fibroblast growth factor receptor eliminates phosphatidylinositol hydrolysis without affecting neuronal differentiation of PC12 cells.
Fibroblast growth factors (FGF) stimulate growth arrest and differentiation in rat pheochromocytoma PC12 cells. We examined the role of phosphatidylinositol (PI) hydrolysis in FGF-induced differentiation of PC12 cells by exploring the biological and biochemical activity of a mutant FGF receptor 1 (flg) defective in stimulation of PI hydrolysis. We show that point mutation at Tyr-766 (Y766F) of the FGF receptor prevents tyrosine phosphorylation of phospholipase C gamma and eliminates acidic FGF (aFGF)-induced stimulation of PI hydrolysis in PC12 cells. Treatment of PC12 cells expressing either wild-type or the Y766F mutant with aFGF led to tyrosine phosphorylation of Shc, the association of Shc with GRB2, a shift in the electrophoretic mobility of the Ras guanine nucleotide-releasing factor, Sos (son of sevenless), and enhancement in mitogen-activated protein kinase phosphorylation. Moreover, stimulation with aFGF led to a typical neurite outgrowth of PC12 cells expressing either wild-type or the Y766F FGF receptor mutant. These experiments indicate that PI hydrolysis is not essential for FGF-induced neuronal differentiation of PC12 cells. Moreover, the aFGF-induced Ras signaling pathway, which is essential for PC12 cell differentiation, is not affected by elimination of PI hydrolysis.
Spivak-Kroizman
T
T
Department of Pharmacology, New York University Medical Center, New York 10016.
Mohammadi
M
M
Hu
P
P
Jaye
M
M
Schlessinger
J
J
Lax
I
I
D000818:Animals; D002454:Cell Differentiation; D016220:Fibroblast Growth Factor 1 / Q000378:metabolism; D000091344:Filaggrin Proteins; D006801:Humans; D006868:Hydrolysis; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D016716:PC12 Cells; D010716:Phosphatidylinositols / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine; D017354:Point Mutation; D051381:Rats; D017468:Receptors, Fibroblast Growth Factor / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis
C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D010716:Phosphatidylinositols; D017468:Receptors, Fibroblast Growth Factor; D011994:Recombinant Proteins; D016220:Fibroblast Growth Factor 1; D019000:Phosphotyrosine; D014443:Tyrosine