Microtubule Attachment and Centromeric Tension Shape the Protein Architecture of the Human Kinetochore

Curr Biol. 2020 Dec 21;30(24):4869-4881.e5. doi: 10.1016/j.cub.2020.09.038. Epub 2020 Oct 8.


The nanoscale protein architecture of the kinetochore plays an integral role in specifying the mechanisms underlying its functions in chromosome segregation. However, defining this architecture in human cells remains challenging because of the large size and compositional complexity of the kinetochore. Here, we use Förster resonance energy transfer to reveal the architecture of individual kinetochore-microtubule attachments in human cells. We find that the microtubule-binding domains of the Ndc80 complex cluster at the microtubule plus end. This clustering occurs only after microtubule attachment, and it increases proportionally with centromeric tension. Surprisingly, Ndc80 complex clustering is independent of the organization and number of its centromeric receptors. Moreover, this clustering is similar in yeast and human kinetochores despite significant differences in their centromeric organizations. These and other data suggest that the microtubule-binding interface of the human kinetochore behaves like a flexible "lawn" despite being nucleated by repeating biochemical subunits.

Keywords: human kinetochore, kinetochore architecture, mitosis, microtubule, cell cycle, centromere, chromosome segregation, FRET.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Centromere / metabolism*
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / isolation & purification
  • Chromosomal Proteins, Non-Histone / metabolism
  • Chromosomal Proteins, Non-Histone / ultrastructure
  • Chromosome Segregation*
  • Cytoskeletal Proteins / genetics
  • Cytoskeletal Proteins / isolation & purification
  • Cytoskeletal Proteins / metabolism
  • Cytoskeletal Proteins / ultrastructure
  • Fluorescence Resonance Energy Transfer
  • Gene Knockdown Techniques
  • Genes, Reporter / genetics
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / genetics
  • HeLa Cells
  • Humans
  • Kinetochores / metabolism
  • Kinetochores / ultrastructure*
  • Microtubules / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Recombinant Proteins / ultrastructure
  • Saccharomyces cerevisiae


  • CENPT protein, human
  • Chromosomal Proteins, Non-Histone
  • Cytoskeletal Proteins
  • NDC80 protein, human
  • Recombinant Proteins
  • centromere protein C
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins