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CDK1-dependent phosphorylation of EZH2 suppresses methylation of H3K27 and promotes osteogenic differentiation of human mesenchymal stem cells

Abstract

Enhancer of zeste homologue 2 (EZH2) is the catalytic subunit of Polycomb repressive complex 2 (PRC2) and catalyses the trimethylation of histone H3 on Lys 27 (H3K27), which represses gene transcription. EZH2 enhances cancer-cell invasiveness and regulates stem cell differentiation. Here, we demonstrate that EZH2 can be phosphorylated at Thr 487 through activation of cyclin-dependent kinase 1 (CDK1). The phosphorylation of EZH2 at Thr 487 disrupted EZH2 binding with the other PRC2 components SUZ12 and EED, and thereby inhibited EZH2 methyltransferase activity, resulting in inhibition of cancer-cell invasion. In human mesenchymal stem cells, activation of CDK1 promoted mesenchymal stem cell differentiation into osteoblasts through phosphorylation of EZH2 at Thr 487. These findings define a signalling link between CDK1 and EZH2 that may have an important role in diverse biological processes, including cancer-cell invasion and osteogenic differentiation of mesenchymal stem cells.

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Figure 1: CDK1 negatively regulates H3K27 trimethylation.
Figure 2: CDK1 interacts with, and phosphorylates, EZH2 at Thr 487.
Figure 3: CDK1-mediated phosphorylation of EZH2 promotes disassociation of EZH2 from SUZ12 and EED and suppresses EZH2 HMTase activity.
Figure 4: Phosphorylation of EZH2 by CDK1 promotes osteogenic differentiation of human mesenchymal stem cells.
Figure 5: CDK1 regulates EZH2-target gene expression in human mesenchymal stem cells.

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Acknowledgements

We thank S. A. Miller and J. Hsu for editorial assistance. We thank A. M. Labaff for characterization of EED antibody. This work was supported by grants from National Institutes of Health (grant R01 CA109311), Kadoorie Charitable Foundations, National Breast Cancer Foundation Inc. and the M. D. Anderson Cancer Center/China Medical University and Hospital Sister Foundation Funds (to M.-C.H), NSC 96-3111-B-039 and NSC 97-3111-B-039 (to M.-C.H., L.-Y.L. and S.-P.Y), NHRI-EX98-9603BC, DOH97-TD-I-111-TM003, DOH98-TD-I-111-TM002 (to L.-Y.L.), DOH97-TD-G-111-041 (to M.-C.H.), DOH99-TD-C-111-005, NSC99-2632-B-039-001-MY3 (to L. –Y.L. and M. –C.H.). In memory of Mrs Serena Lin-Guo for her courageous battle against breast cancer.

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M.-C.H., Y.W. and L.-Y.L. designed the project and wrote the paper. M.-C.H. and L.-Y.L. supervised the research. Y.W. performed most of the experiments in Figs 1, 2, 3. Y.-H.C. performed mesenchymal stem cell experiments in Figs 4 and 5 and experiments investigating the interaction between EZH2 and CDK1. C.-C.L. performed the mass spectrometry analysis. C.-Y.L. generated and characterized the phospho-EZH2 antibody. S.-P.Y. collected and characterized primary human mesenchymal stem cells. J. L., B.S., C. -C.Y. and J.-Y.Y. assisted with experiments. All authors participated in interpreting the results.

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Correspondence to Long-Yuan Li or Mien-Chie Hung.

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The authors declare no competing financial interests.

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Wei, Y., Chen, YH., Li, LY. et al. CDK1-dependent phosphorylation of EZH2 suppresses methylation of H3K27 and promotes osteogenic differentiation of human mesenchymal stem cells. Nat Cell Biol 13, 87–94 (2011). https://doi.org/10.1038/ncb2139

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