Molecular mechanisms facilitating the initial kinetochore encounter with spindle microtubules

J Cell Biol. 2017 Jun 5;216(6):1609-1622. doi: 10.1083/jcb.201608122. Epub 2017 Apr 26.

Abstract

The initial kinetochore (KT) encounter with a spindle microtubule (MT; KT capture) is one of the rate-limiting steps in establishing proper KT-MT interaction during mitosis. KT capture is facilitated by multiple factors, such as MT extension in various directions, KT diffusion, and MT pivoting. In addition, KTs generate short MTs, which subsequently interact with a spindle MT. KT-derived MTs may facilitate KT capture, but their contribution is elusive. In this study, we find that Stu1 recruits Stu2 to budding yeast KTs, which promotes MT generation there. By removing Stu2 specifically from KTs, we show that KT-derived MTs shorten the half-life of noncaptured KTs from 48-49 s to 28-34 s. Using computational simulation, we found that multiple factors facilitate KT capture redundantly or synergistically. In particular, KT-derived MTs play important roles both by making a significant contribution on their own and by synergistically enhancing the effects of KT diffusion and MT pivoting. Our study reveals fundamental mechanisms facilitating the initial KT encounter with spindle MTs.

Publication types

  • Video-Audio Media

MeSH terms

  • Computer Simulation
  • Diffusion
  • Kinetochores / metabolism*
  • Microscopy, Fluorescence
  • Microscopy, Video
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism*
  • Microtubules / metabolism*
  • Mitosis / physiology*
  • Models, Biological
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Spindle Apparatus / metabolism*
  • Time Factors
  • Time-Lapse Imaging

Substances

  • Microtubule-Associated Proteins
  • STU1 protein, S cerevisiae
  • STU2 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins