CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore

Cell. 2008 Dec 12;135(6):1039-52. doi: 10.1016/j.cell.2008.10.019.

Abstract

Kinetochore specification and assembly requires the targeted deposition of specialized nucleosomes containing the histone H3 variant CENP-A at centromeres. However, CENP-A is not sufficient to drive full-kinetochore assembly, and it is not clear how centromeric chromatin is established. Here, we identify CENP-W as a component of the DNA-proximal constitutive centromere-associated network (CCAN) of proteins. We demonstrate that CENP-W forms a DNA-binding complex together with the CCAN component CENP-T. This complex directly associates with nucleosomal DNA and with canonical histone H3, but not with CENP-A, in centromeric regions. CENP-T/CENP-W functions upstream of other CCAN components with the exception of CENP-C, an additional putative DNA-binding protein. Our analysis indicates that CENP-T/CENP-W and CENP-C provide distinct pathways to connect the centromere with outer kinetochore assembly. In total, our results suggest that the CENP-T/CENP-W complex is directly involved in establishment of centromere chromatin structure coordinately with CENP-A.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Autoantigens / genetics
  • Autoantigens / metabolism
  • Centromere Protein A
  • Centromere*
  • Chickens
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism*
  • DNA / metabolism*
  • HeLa Cells
  • Histones / metabolism
  • Humans
  • Kinetochores / metabolism*
  • Mutation
  • Nucleosomes / metabolism

Substances

  • Autoantigens
  • CENPA protein, human
  • CENPT protein, human
  • CENPW protein, human
  • Centromere Protein A
  • Chromosomal Proteins, Non-Histone
  • Histones
  • Nucleosomes
  • DNA