Centromeres are maintained by fastening CENP-A to DNA and directing an arginine anchor-dependent nucleosome transition

Nat Commun. 2017 Jun 9:8:15775. doi: 10.1038/ncomms15775.

Abstract

Maintaining centromere identity relies upon the persistence of the epigenetic mark provided by the histone H3 variant, centromere protein A (CENP-A), but the molecular mechanisms that underlie its remarkable stability remain unclear. Here, we define the contributions of each of the three candidate CENP-A nucleosome-binding domains (two on CENP-C and one on CENP-N) to CENP-A stability using gene replacement and rapid protein degradation. Surprisingly, the most conserved domain, the CENP-C motif, is dispensable. Instead, the stability is conferred by the unfolded central domain of CENP-C and the folded N-terminal domain of CENP-N that becomes rigidified 1,000-fold upon crossbridging CENP-A and its adjacent nucleosomal DNA. Disrupting the 'arginine anchor' on CENP-C for the nucleosomal acidic patch disrupts the CENP-A nucleosome structural transition and removes CENP-A nucleosomes from centromeres. CENP-A nucleosome retention at centromeres requires a core centromeric nucleosome complex where CENP-C clamps down a stable nucleosome conformation and CENP-N fastens CENP-A to the DNA.

Publication types

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

MeSH terms

  • Animals
  • Arginine / metabolism*
  • Centromere / chemistry
  • Centromere / genetics
  • Centromere / metabolism*
  • Centromere Protein A / chemistry
  • Centromere Protein A / genetics
  • Centromere Protein A / metabolism*
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism
  • DNA / genetics
  • DNA / metabolism*
  • Female
  • Humans
  • Male
  • Mice
  • Nucleosomes / chemistry
  • Nucleosomes / genetics
  • Nucleosomes / metabolism*
  • Protein Binding
  • Protein Domains

Substances

  • Centromere Protein A
  • Chromosomal Proteins, Non-Histone
  • Nucleosomes
  • centromere protein C
  • DNA
  • Arginine