title: Undetectable histone O-GlcNAcylation in mammalian cells.
abstract: O-GlcNAcylation is a posttranslational modification catalyzed by the O-Linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) and reversed by O-GlcNAcase (OGA). Numerous transcriptional regulators, including chromatin modifying enzymes, transcription factors, and co-factors, are targeted by O-GlcNAcylation, indicating that this modification is central for chromatin-associated processes. Recently, OGT-mediated O-GlcNAcylation was reported to be a novel histone modification, suggesting a potential role in directly coordinating chromatin structure and function. In contrast, using multiple biochemical approaches, we report here that histone O-GlcNAcylation is undetectable in mammalian cells. Conversely, O-GlcNAcylation of the transcription regulators Host Cell Factor-1 (HCF-1) and Ten-Eleven Translocation protein 2 (TET2) could be readily observed. Our study raises questions on the occurrence and abundance of O-GlcNAcylation as a histone modification in mammalian cells and reveals technical complications regarding the detection of genuine protein O-GlcNAcylation. Therefore, the identification of the specific contexts in which histone O-GlcNAcylation might occur is still to be established.
authors.affiliation: a Maisonneuve-Rosemont Hospital Research Center and Department of Medicine; University of Montréal ; Montréal, Québec , Canada.
authors.forename: Jessica
authors.initials: J
authors.lastname: Gagnon
authors.forename: Salima
authors.initials: S
authors.lastname: Daou
authors.forename: Natalia
authors.initials: N
authors.lastname: Zamorano
authors.forename: Nicholas V G
authors.initials: NV
authors.lastname: Iannantuono
authors.forename: Ian
authors.initials: I
authors.lastname: Hammond-Martel
authors.forename: Nazar
authors.initials: N
authors.lastname: Mashtalir
authors.forename: Eric
authors.initials: E
authors.lastname: Bonneil
authors.forename: Hugo
authors.initials: H
authors.lastname: Wurtele
authors.forename: Pierre
authors.initials: P
authors.lastname: Thibault
authors.forename: El Bachir
authors.initials: el B
authors.lastname: Affar
mesh_terms: D000215:Acylation; D000818:Animals; D002843:Chromatin / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D049308:Dioxygenases; D044127:Epigenesis, Genetic; D006031:Glycosylation; D057809:HEK293 Cells; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D051863:Host Cell Factor C1 / Q000235:genetics*; D006801:Humans; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D001619:beta-N-Acetylhexosaminidases / Q000235:genetics* / Q000378:metabolism
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; C497692:HCFC1 protein, human; D006657:Histones; D051863:Host Cell Factor C1; D011518:Proto-Oncogene Proteins; D049308:Dioxygenases; C541081:TET2 protein, human; D017351:N-Acetylglucosaminyltransferases; C063057:UDP-N-acetylglucosamine-peptide beta-N-acetylglucosaminyltransferase; C110628:hexosaminidase C; D001619:beta-N-Acetylhexosaminidases
keywords: Chromatin; Epigenetics; H2B K120ub, Histone H2B lysine 120 monoubiquitination; H2B S112 O-GlcNAc, Histone H2B serine 112 O-GlcNAc; HCF-1; HCF-1, Host Cell Factor-1; Histone; O-GlcNAc; O-GlcNAc, O-Linked N-acetylglucosamine; O-GlcNAcylation; OGA, O-GlcNAcase; OGT; OGT, O-Linked N-acetylglucosamine transferase; PUGNAc, O-(2-acetamido-2-deoxyglucopyranosylidene) amino N-phenylcarbamate; Polycomb; TET2; TET2, Ten-Eleven Translocation protein 2; UDP-GlcNAc, Uridine Diphosphate N-Acetylglucosamine; WGA, Wheat Germ Agglutinin.; posttranslational modification