Heterochromatin boundaries are hotspots for de novo kinetochore formation

Nat Cell Biol. 2011 Jun 19;13(7):799-808. doi: 10.1038/ncb2272.

Abstract

The centromere-specific histone H3 variant CENH3 (also known as CENP-A) is considered to be an epigenetic mark for establishment and propagation of centromere identity. Pulse induction of CENH3 (Drosophila CID) in Schneider S2 cells leads to its incorporation into non-centromeric regions and generates CID islands that resist clearing from chromosome arms for multiple cell generations. We demonstrate that CID islands represent functional ectopic kinetochores, which are non-randomly distributed on the chromosome and show a preferential localization near telomeres and pericentric heterochromatin in transcriptionally silent, intergenic chromatin domains. Although overexpression of heterochromatin protein 1 (HP1) or increasing histone acetylation interferes with CID island formation on a global scale, induction of a locally defined region of synthetic heterochromatin by targeting HP1-LacI fusions to stably integrated Lac operator arrays produces a proximal hotspot for CID deposition. These data indicate that the characteristics of regions bordering heterochromatin promote de novo kinetochore assembly and thereby contribute to centromere identity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylation
  • Animals
  • Cell Line
  • Centromere Protein A
  • Chromatin Assembly and Disassembly
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism
  • Chromosomes, Insect / metabolism*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Drosophila / genetics
  • Drosophila / metabolism*
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism*
  • Heterochromatin / metabolism*
  • Histones / genetics
  • Histones / metabolism*
  • Kinetochores / metabolism*
  • Lac Operon
  • Lac Repressors / genetics
  • Lac Repressors / metabolism
  • Protein Processing, Post-Translational
  • Recombinant Fusion Proteins / metabolism
  • Telomere / metabolism
  • Time Factors
  • Transfection
  • Up-Regulation

Substances

  • Centromere Protein A
  • Chromosomal Proteins, Non-Histone
  • Cid protein, Drosophila
  • DNA-Binding Proteins
  • Drosophila Proteins
  • Heterochromatin
  • Histones
  • Lac Repressors
  • Recombinant Fusion Proteins
  • heterochromatin-specific nonhistone chromosomal protein HP-1