Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

GlcNAcylation of histone H2B facilitates its monoubiquitination

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.

Your institute does not have access to this article

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

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.

References

  1. Strahl, B. D. & Allis, C. D. The language of covalent histone modifications. Nature 403, 41–45 (2000)

    ADS  CAS  Article  Google Scholar 

  2. Kouzarides, T. Chromatin modifications and their function. Cell 128, 693–705 (2007)

    CAS  Article  Google Scholar 

  3. Li, B., Carey, M. & Workman, J. L. The role of chromatin during transcription. Cell 128, 707–719 (2007)

    CAS  Article  Google Scholar 

  4. Berger, S. L. The complex language of chromatin regulation during transcription. Nature 447, 407–412 (2007)

    ADS  CAS  Article  Google Scholar 

  5. Hart, G. W., Housley, M. P. & Slawson, C. Cycling of O-linked β-N-acetylglucosamine on nucleocytoplasmic proteins. Nature 446, 1017–1022 (2007)

    ADS  CAS  Article  Google Scholar 

  6. Love, D. C. & Hanover, J. A. The hexosamine signaling pathway: deciphering the ‘O-GlcNAc code’. Sci. STKE 2005, re13 (2005)

    PubMed  Google Scholar 

  7. Yang, X., Zhang, F. & Kudlow, J. E. Recruitment of O-GlcNAc transferase to promoters by corepressor mSin3A: coupling protein O-GlcNAcylation to transcriptional repression. Cell 110, 69–80 (2002)

    CAS  Article  Google Scholar 

  8. Gambetta, M. C., Oktaba, K. & Muller, J. Essential role of the glycosyltransferase sxc/Ogt in polycomb repression. Science 325, 93–96 (2009)

    ADS  CAS  Article  Google Scholar 

  9. Fujiki, R. et al. GlcNAcylation of a histone methyltransferase in retinoic-acid-induced granulopoiesis. Nature 459, 455–459 (2009)

    ADS  CAS  Article  Google Scholar 

  10. Wang, Z. et al. Extensive crosstalk between O-GlcNAcylation and phosphorylation regulates cytokinesis. Sci. Signal. 3, ra2 (2010)

    PubMed  PubMed Central  Google Scholar 

  11. Sakabe, K., Wang, Z. & Hart, G. W. β-N-acetylglucosamine (O-GlcNAc) is part of the histone code. Proc. Natl Acad. Sci. USA 107, 19915–19920 (2010)

    ADS  CAS  Article  Google Scholar 

  12. Luger, K. et al. Crystal structure of the nucleosome core particle at 2.8 Å resolution. Nature 389, 251–260 (1997)

    ADS  CAS  Article  Google Scholar 

  13. Das, C., Lucia, M. S., Hansen, K. C. & Tyler, J. K. CBP/p300-mediated acetylation of histone H3 on lysine 56. Nature 459, 113–117 (2009)

    ADS  CAS  Article  Google Scholar 

  14. Dang, W. et al. Histone H4 lysine 16 acetylation regulates cellular lifespan. Nature 459, 802–807 (2009)

    ADS  CAS  Article  Google Scholar 

  15. Dong, L. & Xu, C. W. Carbohydrates induce mono-ubiquitination of H2B in yeast. J. Biol. Chem. 279, 1577–1580 (2004)

    CAS  Article  Google Scholar 

  16. Yoshida, Y. et al. E3 ubiquitin ligase that recognizes sugar chains. Nature 418, 438–442 (2002)

    ADS  CAS  Article  Google Scholar 

  17. Kim, J. et al. RAD6-Mediated transcription-coupled H2B ubiquitylation directly stimulates H3K4 methylation in human cells. Cell 137, 459–471 (2009)

    CAS  Article  Google Scholar 

  18. Dentin, R. et al. Hepatic glucose sensing via the CREB coactivator CRTC2. Science 319, 1402–1405 (2008)

    ADS  CAS  Article  Google Scholar 

  19. Chikanishi, T. et al. Glucose-induced expression of MIP-1 genes requires O-GlcNAc transferase in monocytes. Biochem. Biophys. Res. Commun. 394, 865–870 (2010)

    CAS  Article  Google Scholar 

  20. Jackson, S. P. & Tjian, R. O-glycosylation of eukaryotic transcription factors: implications for mechanisms of transcriptional regulation. Cell 55, 125–133 (1988)

    CAS  Article  Google Scholar 

  21. Sinclair, D. A. et al. Drosophila O-GlcNAc transferase (OGT) is encoded by the Polycomb group (PcG) gene, super sex combs (sxc). Proc. Natl Acad. Sci. USA 106, 13427–13432 (2009)

    ADS  CAS  Article  Google Scholar 

  22. Sawatsubashi, S. et al. A histone chaperone, DEK, transcriptionally coactivates a nuclear receptor. Genes Dev. 24, 159–170 (2009)

    Article  Google Scholar 

  23. Fujiki, R. et al. Ligand-induced transrepression by VDR through association of WSTF with acetylated histones. EMBO J. 24, 3881–3894 (2005)

    CAS  Article  Google Scholar 

  24. He, H. H. et al. Nucleosome dynamics define transcriptional enhancers. Nature Genet. 42, 343–347 (2010)

    CAS  Article  Google Scholar 

  25. Minsky, N. et al. Monoubiquitinated H2B is associated with the transcribed region of highly expressed genes in human cells. Nature Cell Biol. 10, 483–488 (2008)

    CAS  Article  Google Scholar 

Download references

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.).

Author information

Authors and Affiliations

Authors

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.

Ethics declarations

Competing interests

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)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature10656

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing