Abstract
Organization of chromatin by epigenetic mechanisms is essential for establishing and maintaining cellular identity in developing and adult organisms. A key question that remains unresolved about this process is how epigenetic marks are transmitted to the next cell generation during cell division. Here we provide a model to explain how trimethylated Lys 27 of histone 3 (H3K27me3), which is catalysed by the EZH2-containing Polycomb Repressive Complex 2 (PRC2), is maintained in proliferating cells. We show that the PRC2 complex binds to the H3K27me3 mark and colocalizes with this mark in G1 phase and with sites of ongoing DNA replication. Efficient binding requires an intact trimeric PRC2 complex containing EZH2, EED and SUZ12, but is independent of the catalytic SET domain of EZH2. Using a heterologous reporter system, we show that transient recruitment of the PRC2 complex to chromatin, upstream of the transcriptional start site, is sufficient to maintain repression through endogenous PRC2 during subsequent cell divisions. Thus, we suggest that once the H3K27me3 is established, it recruits the PRC2 complex to maintain the mark at sites of DNA replication, leading to methylation of H3K27 on the daughter strands during incorporation of newly synthesized histones. This mechanism ensures maintenance of the H3K27me3 epigenetic mark in proliferating cells, not only during DNA replication when histones synthesized de novo are incorporated, but also outside S phase, thereby preserving chromatin structure and transcriptional programs.
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Change history
28 October 2008
In the version of this article initially published, the order of the labels siRNA cyclophilin B and siRNA SUZ12 in figure 5 and GAL4-EZH2WT and GAL4-EZH2ΔSET in figure 7a were reversed. The corrected panels are shown below. These errors have also been corrected in the HTML and PDF versions of the article.
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Acknowledgements
We thank Danny Reinberg for providing the 5×Gal4TKLucNeo reporter plasmid. We thank members of the Helin laboratory for technical advice and fruitful discussion. This work was supported by grants from the Novo Nordisk Foundation, Association for International Cancer Research, the Danish Cancer Society, the Danish Medical Research Council, the Danish Natural Science Research Council and the Danish National Research Foundation.
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K.H.H. performed most of the experiments; A.P.B. performed ChIP analysis including qPCR on material provided by K.H.H., except for the experiment in Fig. 6c, which was performed by S.S.G.; DP made the ΔSANT2 mutant of EZH2; N.D. helped to establish the Gal4EZH2WT and ΔSET reporter cell lines; A.M. cloned the shRNA construct for Gal4; J.R. performed the final mass spectrometry analysis on samples provided by K.H.H.; ML performed the confocal microscopy analysis on cell stainings provided by K.H.H.; K.H.H. and K.H. jointly conceived the project and wrote the manuscript.
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Hansen, K., Bracken, A., Pasini, D. et al. A model for transmission of the H3K27me3 epigenetic mark. Nat Cell Biol 10, 1291–1300 (2008). https://doi.org/10.1038/ncb1787
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DOI: https://doi.org/10.1038/ncb1787
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