Molecular control of kinetochore-microtubule dynamics and chromosome oscillations

Nat Cell Biol. 2010 Apr;12(4):319-29. doi: 10.1038/ncb2033. Epub 2010 Mar 14.

Abstract

Chromosome segregation in metazoans requires the alignment of sister kinetochores on the metaphase plate. During chromosome alignment, bioriented kinetochores move chromosomes by regulating the plus-end dynamics of the attached microtubules. The bundles of kinetochore-bound microtubules alternate between growth and shrinkage, leading to regular oscillations along the spindle axis. However, the molecular mechanisms that coordinate microtubule plus-end dynamics remain unknown. Here we show that centromere protein (CENP)-H, a subunit of the CENP-A nucleosome-associated and CENP-A distal complexes (CENP-A NAC/CAD), is essential for this coordination, because kinetochores lacking CENP-H establish bioriented attachments but fail to generate regular oscillations, as a result of an uncontrolled rate of microtubule plus-end turnover. These alterations lead to rapid erratic movements that disrupt metaphase plate organization. We also show that the abundance of the CENP-A NAC/CAD subunits CENP-H and CENP-I dynamically change on individual sister kinetochores in vivo, because they preferentially bind the sister kinetochore attached to growing microtubules, and that one other subunit, CENP-Q, binds microtubules in vitro. We therefore propose that CENP-A NAC/CAD is a direct regulator of kinetochore-microtubule dynamics, which physically links centromeric DNA to microtubule plus ends.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autoantigens / metabolism
  • Biosensing Techniques
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism*
  • Centromere Protein A
  • Chromosomal Proteins, Non-Histone / metabolism
  • Chromosome Segregation*
  • Chromosomes, Human / metabolism*
  • DNA-Binding Proteins / metabolism
  • Fluorescence Resonance Energy Transfer
  • HeLa Cells
  • Humans
  • Kinetochores / metabolism*
  • Metaphase* / genetics
  • Microscopy, Confocal
  • Microtubules / genetics
  • Microtubules / metabolism*
  • Protein Binding
  • RNA Interference
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction* / genetics
  • Time Factors
  • Transfection
  • Tubulin / metabolism

Substances

  • Autoantigens
  • CENPA protein, human
  • CENPH protein, human
  • CENPI protein, human
  • Cell Cycle Proteins
  • Centromere Protein A
  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • Recombinant Fusion Proteins
  • Tubulin