Engagement of DNA and H3K27me3 by the CBX8 chromodomain drives chromatin association

Nucleic Acids Res. 2019 Mar 18;47(5):2289-2305. doi: 10.1093/nar/gky1290.

Abstract

Polycomb repressive complex 1 (PRC1) is critical for mediating gene repression during development and adult stem cell maintenance. Five CBX proteins, CBX2,4,6,7,8, form mutually exclusive PRC1 complexes and are thought to play a role in the association of PRC1 with chromatin. Specifically, the N-terminal chromodomain (CD) in the CBX proteins is thought to mediate specific targeting to methylated histones. For CBX8, however, the chromodomain has demonstrated weak affinity and specificity for methylated histones in vitro, leaving doubt as to its role in CBX8 chromatin association. Here, we investigate the function of the CBX8 CD in vitro and in vivo. We find that the CD is in fact a major driver of CBX8 chromatin association and determine that this is driven by both histone and previously unrecognized DNA binding activity. We characterize the structural basis of histone and DNA binding and determine how they integrate on multiple levels. Notably, we find that the chromatin environment is critical in determining the ultimate function of the CD in CBX8 association.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arginine / chemistry
  • Arginine / metabolism
  • Chromatin / genetics
  • Chromatin / metabolism*
  • DNA / chemistry
  • DNA / genetics
  • DNA / metabolism*
  • HEK293 Cells
  • Histones / chemistry*
  • Histones / metabolism*
  • Humans
  • Methylation
  • Models, Molecular
  • Nucleosomes / genetics
  • Nucleosomes / metabolism
  • Polycomb Repressive Complex 1 / chemistry*
  • Polycomb Repressive Complex 1 / metabolism*
  • Protein Binding
  • Protein Domains

Substances

  • CBX8 protein, human
  • Chromatin
  • Histones
  • Nucleosomes
  • DNA
  • Arginine
  • Polycomb Repressive Complex 1