Higher-order protein assembly controls kinetochore formation

Nat Cell Biol. 2024 Jan;26(1):45-56. doi: 10.1038/s41556-023-01313-7. Epub 2024 Jan 2.


To faithfully segregate chromosomes during vertebrate mitosis, kinetochore-microtubule interactions must be restricted to a single site on each chromosome. Prior work on pair-wise kinetochore protein interactions has been unable to identify the mechanisms that prevent outer kinetochore formation in regions with a low density of CENP-A nucleosomes. To investigate the impact of higher-order assembly on kinetochore formation, we generated oligomers of the inner kinetochore protein CENP-T using two distinct, genetically engineered systems in human cells. Although individual CENP-T molecules interact poorly with outer kinetochore proteins, oligomers that mimic centromeric CENP-T density trigger the robust formation of functional, cytoplasmic kinetochore-like particles. Both in cells and in vitro, each molecule of oligomerized CENP-T recruits substantially higher levels of outer kinetochore components than monomeric CENP-T molecules. Our work suggests that the density dependence of CENP-T restricts outer kinetochore recruitment to centromeres, where densely packed CENP-A recruits a high local concentration of inner kinetochore proteins.

MeSH terms

  • Centromere / genetics
  • Centromere / metabolism
  • Centromere Protein A / genetics
  • Chromosomal Proteins, Non-Histone* / genetics
  • Chromosomal Proteins, Non-Histone* / metabolism
  • Humans
  • Kinetochores* / metabolism
  • Mitosis
  • Nucleosomes


  • Centromere Protein A
  • Chromosomal Proteins, Non-Histone
  • Nucleosomes