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GlcNAcylation of histone H2B facilitates its monoubiquitination

Nature volume 480, pages 557–560 (2011)Cite this article

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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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Figure 1: H2B is GlcNAcylated at the C-terminal S112.
Figure 2: H2B S112 GlcNAc is a glucose-responsive modification linked to K120 monoubiquitination (ub).
Figure 3: GlcNAcylation at S112 facilitates ubiquitination at K120 in H2B.
Figure 4: GlcNAcylated H2B is associated with transcribed genes.

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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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Authors and Affiliations

  1. Institute of Molecular and Cellular Biosciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan,

    Ryoji Fujiki, Waka Hashiba, Hiroki Sekine, Atsushi Yokoyama, Toshihiro Chikanishi, Saya Ito, Yuuki Imai, Fumiaki Ohtake, Hirochika Kitagawa & Shigeaki Kato

  2. Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, New York, 10065, New York, USA

    Jaehoon Kim & Robert G. Roeder

  3. Department of Medical Oncology, Dana-Farber Cancer Institute and Harvard Medical School, Boston, 02115, Massachusetts, USA

    Housheng Hansen He & Myles Brown

  4. Division of Cellular and Molecular Toxicology, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan,

    Katsuhide Igarashi & Jun Kanno

  5. ERATO, Japan Science and Technology Agency, Kawaguchi, Saitama 332-0012, Japan,

    Shigeaki Kato

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  1. Ryoji Fujiki
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Contributions

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).

Corresponding author

Correspondence to Shigeaki Kato.

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The authors declare no competing financial interests.

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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