Molecular architecture of human polycomb repressive complex 2

Claudio Ciferri,Alessio Maiolica,Gabriel C Lander,Ruedi Aebersold,Eva Nogales,Franz Herzog

doi:10.7554/elife.00005

Abstract

Polycomb Repressive Complex 2 (PRC2) is essential for gene silencing, establishing transcriptional repression of specific genes by tri-methylating Lysine 27 of histone H3, a process mediated by cofactors such as AEBP2. In spite of its biological importance, little is known about PRC2 architecture and subunit organization. Here, we present the first three-dimensional electron microscopy structure of the human PRC2 complex bound to its cofactor AEBP2. Using a novel internal protein tagging-method, in combination with isotopic chemical cross-linking and mass spectrometry, we have localized all the PRC2 subunits and their functional domains and generated a detailed map of interactions. The position and stabilization effect of AEBP2 suggests an allosteric role of this cofactor in regulating gene silencing. Regions in PRC2 that interact with modified histone tails are localized near the methyltransferase site, suggesting a molecular mechanism for the chromatin-based regulation of PRC2 activity.DOI:http://dx.doi.org/10.7554/eLife.00005.001.

Highlights

The combined antagonistic activities of the Polycomb group (PcG) and the Trithorax group (TrxG) protein complexes contribute to correct homeotic gene expression and accurate cell fate maintenance
The results showed that the class averages were aligned in 3D in a self-consistent fashion, producing a similar complex architecture at the low resolution expected at this stage, irrespective of the Random Conical Tilt (RCT) structure used as a reference
The Trithorax (TrxG) and Polycomb groups (PcG) of protein complexes play a fundamental role in orchestrating the transcriptional activation or silencing of the genome

Summary

Introduction

The combined antagonistic activities of the Polycomb group (PcG) and the Trithorax group (TrxG) protein complexes contribute to correct homeotic gene expression and accurate cell fate maintenance. The PRC2 core, conserved from Drosophila to humans, is composed of four proteins that add up to about 230 kDa (Figure 1A) (see Margueron and Reinberg, 2010 for a recent review): EED (present in different isoforms), either one of the two methyltranferases Ezh or Ezh (Ezh1/2), Suz, and either RbAp46 or RbAp48 (RbAp46/48). Both Ezh and Ezh contain enzymatic methyl-transferase activity within a C-terminal SET (Su(var), Enhancer-of-zeste, Trithorax) domain. Ezh activity appears to depend on interaction with both Suz and the WD40 domain in EED

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Results

Conclusion