title: RING1B O-GlcNAcylation regulates gene targeting of polycomb repressive complex 1 in human embryonic stem cells.
abstract: O-linked-N-acetylglucosamine (O-GlcNAc) post-translationally modifies and regulates thousands of proteins involved in various cellular mechanisms. Recently, O-GlcNAc has been linked to human embryonic stem cells (hESC) differentiation, however the identity and function of O-GlcNAc proteins regulating hESC remain unknown. Here, we firstly identified O-GlcNAc modified human stem cell regulators such as hnRNP K, HP1γ, and especially RING1B/RNF2. Thereafter, we focused our work on RING1B which is the catalytic subunit of the polycomb repressive complex 1 (PRC1) a major epigenetic repressor essential for pluripotency maintenance and differentiation. By point-mutation, we show that T(250)/S(251) and S(278) RING1B residues are bearing O-GlcNAc, and that T(250)/S(251) O-GlcNAcylation decreases during differentiation. O-GlcNAc seems to regulate RING1B-DNA binding as suggested by our ChIP-sequencing results. Non-O-GlcNAcylated RING1B is found to be enriched near cell cycle genes whereas O-GlcNAcylated RING1B seems preferentially enriched near neuronal genes. Our data suggest that during hESC differentiation, the decrease of RING1B O-GlcNAcylation might enable PRC1 to switch its target to induce neuron differentiation. Overall, we demonstrate that O-GlcNAc modifies and regulates an essential epigenetic tool, RING1B, which may contribute to hESC pluripotency maintenance and differentiation.
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01 Centros, 138668 Singapore; Department of Biomedical Engineering, Faculty of Engineering, National University of Singapore, 9 Engineering Drive 1, 117575 Singapore; Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), 61 Biopolis Drive, #08-01 Proteos, 138673 Singapore. Electronic address: jpmaury@imcb.a-star.edu.sg.
authors.forename: Julien Jean Pierre
authors.initials: JJ
authors.lastname: Maury
authors.affiliation: Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), 61 Biopolis Drive, #08-01 Proteos, 138673 Singapore; Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, 117543 Singapore.
authors.forename: Chadi A
authors.initials: CA
authors.lastname: El Farran
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01 Centros, 138668 Singapore.
authors.forename: Daniel
authors.initials: D
authors.lastname: Ng
authors.affiliation: Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), 61 Biopolis Drive, #08-01 Proteos, 138673 Singapore; Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, 117543 Singapore.
authors.forename: Yuin-Han
authors.initials: YH
authors.lastname: Loh
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01 Centros, 138668 Singapore.
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Bi
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01 Centros, 138668 Singapore; Université de Rouen, Normandie Université, Laboratoire Glycobiologie et Matrice Extracellulaire Végétale (Glyco-MEV) EA 4358, IRIB, VASI, Faculté des Sciences et Techniques, Mont-Saint-Aignan Cédex 76821, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Bardor
authors.affiliation: Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01 Centros, 138668 Singapore; Department of Biomedical Engineering, Faculty of Engineering, National University of Singapore, 9 Engineering Drive 1, 117575 Singapore. Electronic address: andre_choo@bti.a-star.edu.sg.
authors.forename: Andre Boon-Hwa
authors.initials: AB
authors.lastname: Choo
mesh_terms: D000595:Amino Acid Sequence; D002454:Cell Differentiation; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D018390:Gene Targeting; D006031:Glycosylation; D034641:Heterogeneous-Nuclear Ribonucleoprotein K / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D063150:Polycomb Repressive Complex 1 / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding
chemical_list: C518294:CBX3 protein, human; D002868:Chromosomal Proteins, Non-Histone; D034641:Heterogeneous-Nuclear Ribonucleoprotein K; D004247:DNA; D063150:Polycomb Repressive Complex 1; C489841:RNF2 protein, human