Abstract
Chromatin reorganization is governed by multiple post-translational modifications of chromosomal proteins and DNA1,2. These histone modifications are reversible, dynamic events that can regulate DNA-driven cellular processes3,4. However, the molecular mechanisms that coordinate histone modification patterns remain largely unknown. In metazoans, reversible protein modification by O-linked N-acetylglucosamine (GlcNAc) is catalysed by two enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA)5,6. However, the significance of GlcNAcylation in chromatin reorganization remains elusive. Here we report that histone H2B is GlcNAcylated at residue S112 by OGT in vitro and in living cells. Histone GlcNAcylation fluctuated in response to extracellular glucose through the hexosamine biosynthesis pathway (HBP)5,6. H2B S112 GlcNAcylation promotes K120 monoubiquitination, in which the GlcNAc moiety can serve as an anchor for a histone H2B ubiquitin ligase. H2B S112 GlcNAc was localized to euchromatic areas on fly polytene chromosomes. In a genome-wide analysis, H2B S112 GlcNAcylation sites were observed widely distributed over chromosomes including transcribed gene loci, with some sites co-localizing with H2B K120 monoubiquitination. These findings suggest that H2B S112 GlcNAcylation is a histone modification that facilitates H2BK120 monoubiquitination, presumably for transcriptional activation.
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Acknowledgements
We thank A. Miyajima, S. Saito and N. Moriyama for experimental support, and M. Yamaki for manuscript preparation. We also thank Y. Maekawa, J. Seta and N. Iwasaki for support with MS. This work was supported in part by The Naito Foundation, the Astellas foundation (to R.F.), the Ministry of Education, Culture, Sports, Science and Technology (MEXT) and the Japan Society for the Promotion of Science (to R.F. and S.K.).
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S.K. planned the study with H.K.; R.G.R. and M.B. provided support and general guidance; R.F. designed the study and performed the experiments with H.S. (α-O-GlcNAc purification), A.Y. (LC–MS/MS), W.H. (O-GlcNAc site mapping), T.C. (in vitro OGT assay), S.I. (Drosophila analysis), Y.I., H.H.H. (ChIP-seq), F.O., J.K. (in vitro monoubiquitination assay), K.I. and J.K (microarray).
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Supplementary information
Supplementary Information
The file contains Supplementary Text, Supplementary Figures 1-25 with legends and legends for Supplementary Table 1-3. (PDF 11414 kb)
Supplementary Table 1
This file shows a list of identified GlcNAcylated proteins in chromatin (Please see Supplementary Information file for full legend.) (XLS 168 kb)
Supplementary Table 2
This file shows microarray analysis of HeLa cells cultured under specific experimental conditions (Please see Supplementary Information file for full legend.) (XLS 6550 kb)
Supplementary Table 3
This file shows a list of genes harboring H2B S112 GlcNAc in the promoter or 50 kbp within the gene body, and the expression levels (Please see Supplementary Information file for full legend.) (XLS 1314 kb)
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Fujiki, R., Hashiba, W., Sekine, H. et al. GlcNAcylation of histone H2B facilitates its monoubiquitination. Nature 480, 557–560 (2011). https://doi.org/10.1038/nature10656
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DOI: https://doi.org/10.1038/nature10656
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