CENP-H-containing complex facilitates centromere deposition of CENP-A in cooperation with FACT and CHD1

Mol Biol Cell. 2009 Sep;20(18):3986-95. doi: 10.1091/mbc.e09-01-0065. Epub 2009 Jul 22.

Abstract

Centromere identity is thought to be determined by epigenetic mechanisms. The centromere-specific histone H3 variant CENP-A plays a central role in specifying the locus where the centromere is constructed. However, the precise mechanisms that target CENP-A to centromeric chromatin are poorly understood. Here, we show that facilitates chromatin transcription (FACT) localizes to centromeres in a CENP-H-containing complex-dependent manner. In conditional mutant cell lines for SSRP1, a subunit of FACT, centromere targeting of newly synthesized CENP-A is severely inhibited. The chromatin remodeling factor CHD1 binds to SSRP1 both in vivo and in vitro and associates with centromeres. The centromeric localization of CHD1 is lost in SSRP1-depleted cells. RNA interference knockdown of CHD1 leads to a decrease in the amount of centromere localized CENP-A. These findings indicate that the CENP-H-containing complex facilitates deposition of newly synthesized CENP-A into centromeric chromatin in cooperation with FACT and CHD1.

MeSH terms

  • Animals
  • Autoantigens / metabolism*
  • Centromere / metabolism*
  • Centromere Protein A
  • Chickens
  • Chromosomal Proteins, Non-Histone / metabolism*
  • DNA Helicases / chemistry
  • DNA Helicases / metabolism*
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / metabolism*
  • HeLa Cells
  • High Mobility Group Proteins / metabolism*
  • Humans
  • Multiprotein Complexes / metabolism*
  • Mutation / genetics
  • Protein Binding
  • Protein Transport
  • Transcriptional Elongation Factors / metabolism*

Substances

  • Autoantigens
  • CENPA protein, human
  • CENPH protein, human
  • Centromere Protein A
  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • High Mobility Group Proteins
  • Multiprotein Complexes
  • SSRP1 protein, human
  • Transcriptional Elongation Factors
  • DNA Helicases
  • CHD1 protein, human