Glycogen Synthase Kinase-3 Interacts with and Phosphorylates Estrogen Receptor α and Is Involved in the Regulation of Receptor Activity

• Published in: The Journal of biological chemistry Vol. 280; no. 38; pp. 33006 - 33014
• Main Authors: Medunjanin, Senad, Hermani, Alexander, De Servi, Barbara, Grisouard, Jean, Rincke, Gabriele, Mayer, Doris
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 23.09.2005
• Subjects: Amino Acid Motifs; Amino Acid Sequence; Animals; Cell Line, Tumor; Cell Proliferation; COS Cells; DNA Mutational Analysis; DNA, Complementary - metabolism; Enzyme Inhibitors - pharmacology; Estrogen Receptor alpha - metabolism; Gene Expression Regulation; Glycogen Synthase Kinase 3 - metabolism; Humans; Immunoprecipitation; Ligands; Lithium Chloride - pharmacology; Luciferases - metabolism; Models, Biological; Molecular Sequence Data; Mutation; Phosphorylation; Plasmids - metabolism; Protein Binding; Protein Structure, Tertiary; Recombinant Proteins - metabolism; Sequence Homology, Amino Acid; Serine - chemistry; Signal Transduction; Subcellular Fractions - metabolism; Transcription, Genetic; Transcriptional Activation
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M506758200

Like other steroid hormone receptors, estrogen receptor-α (ERα) is a substrate for protein kinases, and phosphorylation has profound effects on the function and activity of this receptor. A number of different kinases have been implicated in ERα regulation. In this report we show by mutational analysis and in vitro kinase assays that ERα is a substrate for glycogen synthase kinase-3 (GSK-3) in vitro and is phosphorylated on two sites, the Ser-102, -104, and -106 motif and Ser-118, both located in the N-terminal transcription activation function (AF-1) domain. GSK-3 forms a complex with ERα in vivo as demonstrated by co-immunoprecipitation from cell lysates. The GSK-3 inhibitor lithium chloride was used to determine the role of GSK-3 in phosphorylation of Ser-102, -104, and -106 and Ser-118 in vivo and to explore the role of these serines in the regulation of ERα function. Treatment of cells with lithium chloride resulted in dephosphorylation of Ser-104 and -106 and Ser-118, which suggests these serine residues as targets for GSK-3 in vivo. Our results further suggest that ERα phosphorylation by GSK-3 stabilizes ERα under resting conditions and modulates ERα transcriptional activity upon ligand binding. Inhibition and constitutive activation of GSK-3, both, resulted in inhibition of ERα transcriptional activity, indicating a function of active as well as inactive GSK-3 in ERα regulation. These findings uncover a novel mechanism for the regulation of ERα-mediated estrogen signaling controlled by a dual action of GSK-3.