Metaphase kinetochore movements are regulated by kinesin-8 motors and microtubule dynamic instability

Mol Biol Cell. 2018 Jun 1;29(11):1332-1345. doi: 10.1091/mbc.E17-11-0667. Epub 2018 Apr 5.

Abstract

During metaphase, sister chromatids are connected to microtubules extending from the opposite spindle poles via kinetochores to protein complexes on the chromosome. Kinetochores congress to the equatorial plane of the spindle and oscillate around it, with kinesin-8 motors restricting these movements. Yet, the physical mechanism underlying kinetochore movements is unclear. We show that kinetochore movements in the fission yeast Schizosaccharomyces pombe are regulated by kinesin-8-promoted microtubule catastrophe, force-induced rescue, and microtubule dynamic instability. A candidate screen showed that among the selected motors only kinesin-8 motors Klp5/Klp6 are required for kinetochore centering. Kinesin-8 accumulates at the end of microtubules, where it promotes catastrophe. Laser ablation of the spindle resulted in kinetochore movement toward the intact spindle pole in wild-type and klp5Δ cells, suggesting that kinetochore movement is driven by pulling forces. Our theoretical model with Langevin description of microtubule dynamic instability shows that kinesin-8 motors are required for kinetochore centering, whereas sensitivity of rescue to force is necessary for the generation of oscillations. We found that irregular kinetochore movements occur for a broader range of parameters than regular oscillations. Thus, our work provides an explanation for how regulation of microtubule dynamic instability contributes to kinetochore congression and the accompanying movements around the spindle center.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromosomes, Fungal / metabolism
  • Hydroxyurea / pharmacology
  • Kinesins / metabolism*
  • Kinetochores / drug effects
  • Kinetochores / metabolism*
  • Metaphase*
  • Microtubule-Associated Proteins / metabolism*
  • Microtubules / drug effects
  • Microtubules / metabolism*
  • Models, Biological
  • Movement
  • Mutation / genetics
  • Schizosaccharomyces / cytology*
  • Schizosaccharomyces / metabolism*
  • Schizosaccharomyces pombe Proteins / metabolism*
  • Spindle Apparatus / drug effects
  • Spindle Apparatus / metabolism

Substances

  • Microtubule-Associated Proteins
  • Schizosaccharomyces pombe Proteins
  • Klp5 protein, S pombe
  • Klp6 protein, S pombe
  • Kinesins
  • Hydroxyurea