title: O-GlcNAcylation stabilizes β-catenin through direct competition with phosphorylation at threonine 41.
abstract: Dysfunctions in Wnt signaling increase β-catenin stability and are associated with cancers, including colorectal cancer. In addition, β-catenin degradation is decreased by nutrient-dependent O-GlcNAcylation. Human colon tumors and colons from mice fed high-carbohydrate diets exhibited higher amounts of β-catenin and O-GlcNAc relative to healthy tissues and mice fed a standard diet, respectively. Administration of the O-GlcNAcase inhibitor thiamet G to mice also increased colonic expression of β-catenin. By ETD-MS/MS, we identified 4 O-GlcNAcylation sites at the N terminus of β-catenin (S23/T40/T41/T112). Furthermore, mutation of serine and threonine residues within the D box of β-catenin reduced O-GlcNAcylation by 75%. Interestingly, elevating O-GlcNAcylation in human colon cell lines drastically reduced phosphorylation at T41, a key residue of the D box responsible for β-catenin stability. Analyses of β-catenin O-GlcNAcylation mutants reinforced T41 as the most crucial residue that controls the β-catenin degradation rate. Finally, inhibiting O-GlcNAcylation decreased the β-catenin/α-catenin interaction necessary for mucosa integrity, whereas O-GlcNAcase silencing improved this interaction. These results suggest that O-GlcNAcylation regulates not only the stability of β-catenin, but also affects its localization at the level of adherens junctions. Accordingly, we propose that O-GlcNAcylation of β-catenin is a missing link between the glucose metabolism deregulation observed in metabolic disorders and the development of cancer.
authors.affiliation: Unit of Structural and Functional Glycobiology, Institut Fédératif de Recherche (IFR) 147, Centre National de la Recherche Scientifique-Unité Mixte de Recherche (CNRS-UMR) 8576, University of Lille 1, Villeneuve d'Ascq, France;
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Olivier-Van Stichelen
authors.affiliation: Unit of Structural and Functional Glycobiology, Institut Fédératif de Recherche (IFR) 147, Centre National de la Recherche Scientifique-Unité Mixte de Recherche (CNRS-UMR) 8576, University of Lille 1, Villeneuve d'Ascq, France;
authors.forename: Vanessa
authors.initials: V
authors.lastname: Dehennaut
authors.affiliation: Exploratory Unit/Systems Biology, Sanofi-Aventis Research and Development, Toulouse, France; and.
authors.forename: Armelle
authors.initials: A
authors.lastname: Buzy
authors.affiliation: Exploratory Unit/Systems Biology, Sanofi-Aventis Research and Development, Toulouse, France; and.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Zachayus
authors.affiliation: Unit of Structural and Functional Glycobiology, Institut Fédératif de Recherche (IFR) 147, Centre National de la Recherche Scientifique-Unité Mixte de Recherche (CNRS-UMR) 8576, University of Lille 1, Villeneuve d'Ascq, France;
authors.forename: Céline
authors.initials: C
authors.lastname: Guinez
authors.affiliation: Unit of Structural and Functional Glycobiology, Institut Fédératif de Recherche (IFR) 147, Centre National de la Recherche Scientifique-Unité Mixte de Recherche (CNRS-UMR) 8576, University of Lille 1, Villeneuve d'Ascq, France;
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Mir
authors.affiliation: Unit of Structural and Functional Glycobiology, Institut Fédératif de Recherche (IFR) 147, Centre National de la Recherche Scientifique-Unité Mixte de Recherche (CNRS-UMR) 8576, University of Lille 1, Villeneuve d'Ascq, France;
authors.forename: Ikram
authors.initials: I
authors.lastname: El Yazidi-Belkoura
authors.affiliation: Tumor Bank, Regional Reference Center in Cancer, Centre Hospitalier Régional Universitaire de Lille (CHRU), University of Lille 2, Lille, France.
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Copin
authors.affiliation: Exploratory Unit/Systems Biology, Sanofi-Aventis Research and Development, Toulouse, France; and.
authors.forename: Didier
authors.initials: D
authors.lastname: Boureme
authors.affiliation: Exploratory Unit/Systems Biology, Sanofi-Aventis Research and Development, Toulouse, France; and.
authors.forename: Denis
authors.initials: D
authors.lastname: Loyaux
authors.affiliation: Exploratory Unit/Systems Biology, Sanofi-Aventis Research and Development, Toulouse, France; and.
authors.forename: Pascual
authors.initials: P
authors.lastname: Ferrara
authors.affiliation: Unit of Structural and Functional Glycobiology, Institut Fédératif de Recherche (IFR) 147, Centre National de la Recherche Scientifique-Unité Mixte de Recherche (CNRS-UMR) 8576, University of Lille 1, Villeneuve d'Ascq, France; tony.lefebvre@univ-lille1.fr.
authors.forename: Tony
authors.initials: T
authors.lastname: Lefebvre
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism*; D000230:Adenocarcinoma / Q000209:etiology / Q000378:metabolism; D022005:Adherens Junctions / Q000378:metabolism / Q000473:pathology; D000595:Amino Acid Sequence; D000818:Animals; D003106:Colon / Q000378:metabolism; D015179:Colorectal Neoplasms / Q000209:etiology / Q000378:metabolism; D004040:Dietary Carbohydrates / Q000378:metabolism / Q000633:toxicity; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D006943:Hyperglycemia / Q000150:complications / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism; D061986:MCF-7 Cells; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000037:antagonists & inhibitors / Q000502:physiology; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D025941:Protein Interaction Mapping; D011499:Protein Processing, Post-Translational; D055550:Protein Stability; D059748:Proteolysis; D034741:RNA, Small Interfering / Q000494:pharmacology; D013912:Threonine / Q000737:chemistry*; D060449:Wnt Signaling Pathway; D051178:alpha Catenin / Q000378:metabolism; D051176:beta Catenin / Q000737:chemistry* / Q000378:metabolism; D001619:beta-N-Acetylhexosaminidases / Q000037:antagonists & inhibitors / Q000502:physiology
chemical_list: D004040:Dietary Carbohydrates; D004791:Enzyme Inhibitors; D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; D051178:alpha Catenin; D051176:beta Catenin; D013912:Threonine; D017351:N-Acetylglucosaminyltransferases; C449880:O-GlcNAc transferase; C110628:hexosaminidase C; D001619:beta-N-Acetylhexosaminidases; D005947:Glucose; D000117:Acetylglucosamine
keywords: ETD-MS/MS; Wnt signaling; cancer
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