Phosphorylation of Glycogen Synthase Kinase-3 and Stimulation of T-cell Factor Signaling following Activation of EP2 and EP4 Prostanoid Receptors by Prostaglandin E2

Recently we have shown that the FPB prostanoid receptor, a G-protein-coupled receptor that couples to Gαq, activates T-cell factor (Tcf)/lymphoid enhancer factor (Lef)-mediated transcriptional activation (Fujino, H., and Regan, J. W. (2001) J. Biol. Chem. 276, 12489–12492). We now report that the EP...

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Published in:	The Journal of biological chemistry Vol. 277; no. 4; pp. 2614 - 2619
Main Authors:	Fujino, Hiromichi, West, Kimberly A., Regan, John W.
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 25-01-2002 American Society for Biochemistry and Molecular Biology
Subjects:	Androstadienes - pharmacology Binding, Competitive Blotting, Western Calcium-Calmodulin-Dependent Protein Kinases - metabolism Cell Differentiation Cell Division Cell Line Cyclic AMP-Dependent Protein Kinases - metabolism Dinoprostone - metabolism DNA-Binding Proteins - metabolism Dose-Response Relationship, Drug Enzyme Inhibitors - pharmacology Genes, Reporter Glycogen Synthase Kinase 3 Glycogen Synthase Kinases Humans Isoquinolines - pharmacology Luciferases - metabolism Lymphoid Enhancer-Binding Factor 1 Phosphatidylinositol 3-Kinases - metabolism Phosphorylation Protein Binding Radioligand Assay Receptors, Prostaglandin E - metabolism Receptors, Prostaglandin E, EP2 Subtype Receptors, Prostaglandin E, EP4 Subtype Signal Transduction Sulfonamides Time Factors Transcription Factors - metabolism Transcription, Genetic Transfection Wortmannin
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Summary:	Recently we have shown that the FPB prostanoid receptor, a G-protein-coupled receptor that couples to Gαq, activates T-cell factor (Tcf)/lymphoid enhancer factor (Lef)-mediated transcriptional activation (Fujino, H., and Regan, J. W. (2001) J. Biol. Chem. 276, 12489–12492). We now report that the EP2 and EP4 prostanoid receptors, which couple to Gαs, also activate Tcf/Lef signaling. By using a Tcf/Lef-responsive luciferase reporter gene, transcriptional activity was stimulated ∼10-fold over basal by 1 h of treatment with prostaglandin E2 (PGE2) in HEK cells that were stably transfected with the human EP2 and EP4receptors. This stimulation of reporter gene activity was accompanied by a PGE2-dependent increase in the phosphorylation of both glycogen synthase kinase-3 (GSK-3) and Akt kinase. H-89, an inhibitor of protein kinase A (PKA), completely blocked the agonist-dependent phosphorylation of GSK-3 in both EP2- and EP4-expressing cells. However, H-89 pretreatment only blocked PGE2-stimulated Lef/Tcf reporter gene activity by 20% in EP4-expressing cells compared with 65% inhibition in EP2-expressing cells. On the other hand wortmannin, an inhibitor of phosphatidylinositol 3-kinase, had the opposite effect and inhibited PGE2-stimulated reporter gene activity to a much greater extent in EP4-expressing cells as compared with EP2-expressing cells. These findings indicate that the activation of Tcf/Lef signaling by EP2 receptors occurs primarily through a PKA-dependent pathway, whereas EP4 receptors activate Tcf/Lef signaling mainly through a phosphatidylinositol 3-kinase-dependent pathway. This is the first indication of a fundamental difference in the signaling potential of EP2 and EP4 prostanoid receptors.
ISSN:	0021-9258 1083-351X
DOI:	10.1074/jbc.M109440200