How the kinetochore couples microtubule force and centromere stretch to move chromosomes

Nat Cell Biol. 2016 Apr;18(4):382-92. doi: 10.1038/ncb3323. Epub 2016 Mar 14.

Abstract

The Ndc80 complex (Ndc80, Nuf2, Spc24 and Spc25) is a highly conserved kinetochore protein essential for end-on anchorage to spindle microtubule plus ends and for force generation coupled to plus-end polymerization and depolymerization. Spc24/Spc25 at one end of the Ndc80 complex binds the kinetochore. The N-terminal tail and CH domains of Ndc80 bind microtubules, and an internal domain binds microtubule-associated proteins (MAPs) such as the Dam1 complex. To determine how the microtubule- and MAP-binding domains of Ndc80 contribute to force production at the kinetochore in budding yeast, we have inserted a FRET tension sensor into the Ndc80 protein about halfway between its microtubule-binding and internal loop domains. The data support a mechanical model of force generation at metaphase where the position of the kinetochore relative to the microtubule plus end reflects the relative strengths of microtubule depolymerization, centromere stretch and microtubule-binding interactions with the Ndc80 and Dam1 complexes.

Publication types

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

MeSH terms

  • Binding Sites / genetics
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Centromere / metabolism*
  • Chromosomes, Fungal / metabolism*
  • Fluorescence Resonance Energy Transfer
  • Kinetics
  • Kinetochores / metabolism*
  • Luminescent Proteins* / genetics
  • Luminescent Proteins* / metabolism
  • Microscopy, Fluorescence
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism
  • Microtubules / metabolism*
  • Models, Biological
  • Mutation
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Protein Binding
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Saccharomycetales / genetics
  • Saccharomycetales / metabolism
  • Time-Lapse Imaging

Substances

  • Cell Cycle Proteins
  • DAM1 protein, S cerevisiae
  • Luminescent Proteins
  • Microtubule-Associated Proteins
  • NDC80 protein, S cerevisiae
  • Nuclear Proteins
  • Saccharomyces cerevisiae Proteins