title: TET3-OGT interaction increases the stability and the presence of OGT in chromatin.
abstract: Gene expression is controlled by alterations in the epigenome, including DNA methylation and histone modification. Recently, it was reported that 5-methylcytosine (5mC) is converted to 5-hydroxymethylcytosine (5hmC) by proteins in the ten-eleven translocation (TET) family. This conversion is believed to be part of the mechanism by which methylated DNA is demethylated. Moreover, histones undergo modifications such as phosphorylation and acetylation. In addition, modification with O-linked-N-acetylglucosamine (O-GlcNAc) by O-GlcNAc transferase (OGT) was recently identified as a novel histone modification. Herein, we focused on TET3, the regulation of which is still unclear. We attempted to elucidate the mechanism of its regulation by biochemical approaches. First, we conducted mass spectrometric analysis in combination with affinity purification of FLAG-TET3, which identified OGT as an important partner of TET3. Co-immunoprecipitation assays using a series of deletion mutants showed that the C-terminal H domain of TET3 was required for its interaction with OGT. Furthermore, we showed that TET3 is GlcNAcylated by OGT, although the GlcNAcylation did not affect the global hydroxylation of methylcytosine by TET3. Moreover, we showed that TET3 enhanced its localization to chromatin through the stabilization of OGT protein. Taken together, we showed a novel function of TET3 that likely supports the function of OGT.
authors.affiliation: Department of Molecular Endocrinology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Ito
authors.forename: Shogo
authors.initials: S
authors.lastname: Katsura
authors.forename: Hiroki
authors.initials: H
authors.lastname: Shimada
authors.forename: Hikaru
authors.initials: H
authors.lastname: Tsuchiya
authors.forename: Masashi
authors.initials: M
authors.lastname: Hada
authors.forename: Tomoko
authors.initials: T
authors.lastname: Okumura
authors.forename: Akira
authors.initials: A
authors.lastname: Sugawara
authors.forename: Atsushi
authors.initials: A
authors.lastname: Yokoyama
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002843:Chromatin / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D049308:Dioxygenases / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D051379:Mice; D017351:N-Acetylglucosaminyltransferases / Q000737:chemistry* / Q000378:metabolism; D055550:Protein Stability; D011518:Proto-Oncogene Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; C578792:TET3 protein, human; D049308:Dioxygenases; C551915:Tet3 protein, mouse; D017351:N-Acetylglucosaminyltransferases; C449880:O-GlcNAc transferase