Insights into Centromere DNA Bending Revealed by the Cryo-EM Structure of the Core Centromere Binding Factor 3 with Ndc10

Cell Rep. 2018 Jul 17;24(3):744-754. doi: 10.1016/j.celrep.2018.06.068.

Abstract

The centromere binding factor 3 (CBF3) complex binds the third centromere DNA element in organisms with point centromeres, such as S. cerevisiae. It is an essential complex for assembly of the kinetochore in these organisms, as it facilitates genetic centromere specification and allows association of all other kinetochore components. We determined high-resolution structures of the core complex of CBF3 alone and in association with a monomeric construct of Ndc10, using cryoelectron microscopy (cryo-EM). We identify the DNA-binding site of the complex and present a model in which CBF3 induces a tight bend in centromeric DNA, thus facilitating assembly of the centromeric nucleosome.

Keywords: CBF3; Ndc10; cryo-EM; kinetochore; point centromere.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding Sites
  • Centromere / metabolism*
  • Cryoelectron Microscopy*
  • DNA, Fungal / chemistry*
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / ultrastructure*
  • Kinetochores / chemistry
  • Kinetochores / ultrastructure*
  • Models, Molecular
  • Mutant Proteins / chemistry
  • Mutant Proteins / metabolism
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / ultrastructure*

Substances

  • CBF2 protein, S cerevisiae
  • CBF3b protein, S cerevisiae
  • DNA, Fungal
  • DNA-Binding Proteins
  • Mutant Proteins
  • Saccharomyces cerevisiae Proteins