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Transcriptional repression mediated by the human polycomb-group protein EED involves histone deacetylation

Nature Genetics volume 23, pages 474–478 (1999)Cite this article

Abstract

Polycomb-group (PcG) proteins form multimeric protein complexes, which are involved in maintaining the transcriptional repressive state of genes over successive cell generations1,2,3. Components of PcG complexes and their mutual interactions have been identified and analysed through extensive genetic and biochemical analyses. Molecular mechanisms underlying PcG-mediated repression of gene activity, however, have remained largely unknown. Previously we reported the existence of two distinct human PcG protein complexes4. The EED/EZH protein complex contains the embryonic ectoderm development4,5,6,7,8 (EED) and enhancer of zeste 2 (EZH2; refs 9,10) PcG proteins. The HPC/HPH PcG complex contains the human polycomb 2 (HPC2; ref. 11), human polyhomeotic12 (HPH), BMI1 (ref. 13 ) and RING1 (refs 14,15) proteins. Here we show that EED (refs 4, 5, 6, 7, 8) interacts, both in vitro and in vivo, with histone deacetylase (HDAC) proteins16,17. This interaction is highly specific because the HDAC proteins do not interact with other vertebrate PcG proteins. We further find that histone deacetylation activity co-immunoprecipitates with the EED protein. Finally, the histone deacetylase inhibitor trichostatin A (ref. 17) relieves transcriptional repression mediated by EED, but not by HPC2, a human homologue of polycomb11. Our data indicate that PcG-mediated repression of gene activity involves histone deacetylation. This mechanistic link between two distinct, global gene repression systems is accomplished through the interaction of HDAC proteins with a particular PcG protein, EED.

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Figure 1: Two-hybrid interactions between the PcG protein EED and HDAC2.
Figure 2: Interactions between EED, EZH2, HDAC1 and HDAC2.
Figure 3: Co-immunoprecipitation of EED, EZH2 and histone deacetylase activity from human 293 cells.
Figure 4: Incubation with the histone deacetylase inhibitor TSA results in relief of EED- and HDAC2-mediated repression, but not of HPC2-mediated repression.

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References

  1. Pirrotta, V. PcG complexes and chromatin silencing. Curr. Opin. Genet. Dev. 7, 249–258 ( 1997).

    Article  CAS  Google Scholar 

  2. Kingston, R.E., Bunker, C.A. & Imbalzano, A.N. Repression and activation by multiprotein complexes that alter chromatin structure. Genes Dev. 10, 905–920 (1996).

    Article  CAS  Google Scholar 

  3. Schumacher, A. & Magnuson, T. Murine Polycomb- and trithorax-group genes regulate homeotic pathways and beyond. Trends Genet. 13, 167–170 (1997).

    Article  CAS  Google Scholar 

  4. Sewalt, R.G.A.B. et al. Characterization of interactions between the mammalian Polycomb-group proteins Enx1/EZH2 and EED suggests the existence of different mammalian Polycomb-group protein complexes. Mol. Cell. Biol. 18, 3586–3595 (1998).

    Article  CAS  Google Scholar 

  5. Schumacher, A., Faust, C. & Magnuson, T. Positional cloning of a global regulator of anterior-posterior patterning in mice. Nature 383, 250– 253 (1996).

    Article  CAS  Google Scholar 

  6. Van Lohuizen, M. et al. Interaction of mouse polycomb-group (Pc-G) proteins Enx1 and Enx2 with Eed: indication for seperate Pc-G complexes. Mol. Cell. Biol. 18, 3572–3579 (1998).

    Article  CAS  Google Scholar 

  7. Denisenko, O.N. & Bomsztyk, K. The product of the murine homolog of the Drosophila extra sex combs gene displays transcriptional repressor activity. Mol. Cell. Biol. 17, 4707–4717 (1997).

    Article  CAS  Google Scholar 

  8. Denisenko, O.N. et al. Point mutations in the WD40 domain of Eed block its interaction with Ezh2. Mol. Cell. Biol. 18, 5634– 5642 (1998).

    Article  CAS  Google Scholar 

  9. Hobert, O., Jalla, B. & Ullrich, A. Interaction of Vav with ENX-1, a putative transcriptional regulator of homeobox gene expression. Mol. Cell. Biol. 16, 3066–3073 (1996).

    Article  CAS  Google Scholar 

  10. Laible, G. et al. Mammalian homologues of the Polycomb-group gene Enhancer of zeste mediate gene silencing in Drosophila heterochromatin and at S. cerevisiae telomeres. EMBO J. 16, 3219–3232 (1997).

    Article  CAS  Google Scholar 

  11. Satijn, D.P.E. et al. Interference with the expression of a novel human Polycomb protein, HPC2, results in cellular transformation and apoptosis. Mol. Cell. Biol. 17, 6076–6086 (1997).

    Article  CAS  Google Scholar 

  12. Gunster, M.J. et al. Identification and characterization of interactions between the vertebrate Polycomb-group protein BMI1 and human homologs of Polyhomeotic. Mol. Cell. Biol. 17, 2326– 2335 (1997).

    Article  CAS  Google Scholar 

  13. Van Lohuizen, M. et al. Identification of cooperating oncogenes in E(mu)-myc transgenic mice by provirus tagging. Cell 65 , 737–752 (1991).

    Article  CAS  Google Scholar 

  14. Satijn, D.P.E. et al. RING1 is associated with the Polycomb-group protein complex and acts as a transcriptional repressor. Mol. Cell. Biol. 17, 4105–4113 (1997).

    Article  CAS  Google Scholar 

  15. Satijn, D.P.E. & Otte, A.P. RING1 interacts with multiple Polycomb-group proteins and displays tumorigenic activity. Mol. Cell. Biol. 19, 57–68 (1999).

    Article  CAS  Google Scholar 

  16. Yang, W.-M. et al. Isolation and characterization of cDNAs corresponding to an additional member of the human histone deacetylation gene family. J. Biol. Chem. 272, 28001–28007 (1997).

    Article  CAS  Google Scholar 

  17. Taunton, J., Hassig, C.A. & Schreiber, S.L. A mammalian histone deacetylase related the yeast transcriptional regulator Rpd3p. Science 272, 408–411 (1996).

    Article  CAS  Google Scholar 

  18. Levine, M.A. et al. Molecular cloning of β 3 subunit, a third form of the G protein RT β-subunit polypeptide. Proc. Natl Acad. Sci. USA 87, 2329–2333 (1990).

    Article  CAS  Google Scholar 

  19. Hassig, C.A. et al. A role for histone deacetylase activity in HDAC1-mediated transcriptional repression. Proc. Natl Acad. Sci. USA 95, 3519–3524 (1998).

    Article  CAS  Google Scholar 

  20. Smith, D.B. & Johnson, K.S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67, 31–40 (1988).

    Article  CAS  Google Scholar 

  21. Shao, Z. et al. Stabilization of chromatin structure by PCR1, a Polycomb complex. Cell 98, 37–46 (1999).

    Article  CAS  Google Scholar 

  22. Mizzen, C.A. et al. Histone acetyltransferases: preparation of substrates and assay procedures. Methods Enzymol. 304, 675–696 (1999).

    Article  CAS  Google Scholar 

  23. Turner, B.M. & Fellows, G. Specific antibodies reveal ordered and cell-cycle-related use of histone H4 acetylation sites in mammalian cells. Eur. J. Biochem. 179, 131– 139 (1989).

    Article  CAS  Google Scholar 

  24. Nan, X. et al. Transcriptional repression by the methyl-CpG-binding protein MeCP2 involves a histone deacetylase complex. Nature 393, 386–389 (1998).

    Article  CAS  Google Scholar 

  25. Strouboulis, J. et al. Transcriptional repression by XPc1, a new Polycomb homolog in Xenopus laevis embryos, is independent of histone deacetylase. Mol. Cell. Biol. 19, 3958–3968 (1999).

    Article  CAS  Google Scholar 

  26. Struhl, G. & Brower, D. Early role of the esc+ gene product in the determination of segments in Drosophila. Cell 31, 285–292 (1982).

    Article  CAS  Google Scholar 

  27. Gutjahr, T. et al. The Polycomb-group gene, extra sex combs, encodes a nuclear member of the WD-40 repeat family. EMBO J. 14, 4296–4306 (1995).

    Article  CAS  Google Scholar 

  28. Lai, J.-S. & Herr, W. Ethidium bromide provides a simple test for identifying genuine DNA-independent protein associations. Proc. Natl Acad. Sci. USA 89, 6958– 6962 (1992).

    Article  CAS  Google Scholar 

  29. Monahan, S.J. et al. Interaction between the herpes simplex virus type I origin-binding and DNA polymerase accessory proteins. Virology 241 , 122–130 (1998).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank B. Turner for antibodies against acetylated histone H4; W. Yang and E. Seto for the HDAC cDNAs; M. Hoefkens for help with the two-hybrid analysis, M. Gunster for help with isolation of the EED/EZH complex; R. Sewalt for help with the preparation of the nucleosomes; K. Hamer for help with the GST-fusion proteins; and R. van Driel and D. Satijn for comments on the manuscript.

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  1. E.C. Slater Instituut, BioCentrum Amsterdam, University of Amsterdam, Amsterdam, The Netherlands

    Johan van der Vlag & Arie P. Otte

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Correspondence to Arie P. Otte.

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van der Vlag, J., Otte, A. Transcriptional repression mediated by the human polycomb-group protein EED involves histone deacetylation. Nat Genet 23, 474–478 (1999). https://doi.org/10.1038/70602

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