title: Phosphorylation of TET proteins is regulated via O-GlcNAcylation by the O-linked N-acetylglucosamine transferase (OGT).
abstract: TET proteins oxidize 5-methylcytosine to 5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxylcytosine and thus provide a possible means for active DNA demethylation in mammals. Although their catalytic mechanism is well characterized and the catalytic dioxygenase domain is highly conserved, the function of the regulatory regions (the N terminus and the low-complexity insert between the two parts of the dioxygenase domains) is only poorly understood. Here, we demonstrate that TET proteins are subject to a variety of post-translational modifications that mostly occur at these regulatory regions. We mapped TET modification sites at amino acid resolution and show for the first time that TET1, TET2, and TET3 are highly phosphorylated. The O-linked GlcNAc transferase, which we identified as a strong interactor with all three TET proteins, catalyzes the addition of a GlcNAc group to serine and threonine residues of TET proteins and thereby decreases both the number of phosphorylation sites and site occupancy. Interestingly, the different TET proteins display unique post-translational modification patterns, and some modifications occur in distinct combinations. In summary, our results provide a novel potential mechanism for TET protein regulation based on a dynamic interplay of phosphorylation and O-GlcNAcylation at the N terminus and the low-complexity insert region. Our data suggest strong cross-talk between the modification sites that could allow rapid adaption of TET protein localization, activity, or targeting due to changing environmental conditions as well as in response to external stimuli.
authors.affiliation: Biocenter, Ludwig-Maximilians University Munich, 82152 Planegg-Martinsried.
authors.forename: Christina
authors.initials: C
authors.lastname: Bauer
authors.affiliation: Biocenter, Ludwig-Maximilians University Munich, 82152 Planegg-Martinsried.
authors.forename: Klaus
authors.initials: K
authors.lastname: Göbel
authors.affiliation: Max Planck Institute of Biochemistry, D-82152 Martinsried.
authors.forename: Nagarjuna
authors.initials: N
authors.lastname: Nagaraj
authors.affiliation: Biocenter, Ludwig-Maximilians University Munich, 82152 Planegg-Martinsried.
authors.forename: Christian
authors.initials: C
authors.lastname: Colantuoni
authors.affiliation: Biocenter, Ludwig-Maximilians University Munich, 82152 Planegg-Martinsried.
authors.forename: Mengxi
authors.initials: M
authors.lastname: Wang
authors.affiliation: Biocenter, Ludwig-Maximilians University Munich, 82152 Planegg-Martinsried.
authors.forename: Udo
authors.initials: U
authors.lastname: Müller
authors.affiliation: Institute for Molecular Immunology, Helmholtz Center Munich, 81377 München-Groβhadern.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Kremmer
authors.affiliation: Biocenter, Ludwig-Maximilians University Munich, 82152 Planegg-Martinsried,. Electronic address: a.rottach@lmu.de.
authors.forename: Andrea
authors.initials: A
authors.lastname: Rottach
authors.affiliation: Biocenter, Ludwig-Maximilians University Munich, 82152 Planegg-Martinsried,; Center for Integrated Protein Science Munich (CIPSM), 81377 München, Germany. Electronic address: h.leonhardt@lmu.de.
authors.forename: Heinrich
authors.initials: H
authors.lastname: Leonhardt
mesh_terms: D000117:Acetylglucosamine; D000215:Acylation / Q000502:physiology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D049308:Dioxygenases / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D006899:Mixed Function Oxygenases; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation / Q000502:physiology; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000502:physiology; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; D006899:Mixed Function Oxygenases; C462539:TET1 protein, human; C578792:TET3 protein, human; D049308:Dioxygenases; C541081:TET2 protein, human; D017351:N-Acetylglucosaminyltransferases; C063057:UDP-N-acetylglucosamine-peptide beta-N-acetylglucosaminyltransferase; D000117:Acetylglucosamine
keywords: 5-Hydroxymethylcytosine (5-hmC); Dioxygenase; Epigenetics; O-Linked N-Acetylglucosamine (O-GlcNAc); OGT; Phosphorylation; Post-translational Modification (PTM); TET Proteins