Suzuki, AussieBadger, Benjamin LHaase, JulianOhashi, TomooErickson, Harold PSalmon, Edward DBloom, Kerry2018-04-012018-04-012016-041465-73921476-4679https://hdl.handle.net/10161/16464The 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.Chromosomes, FungalCentromereKinetochoresMicrotubulesSaccharomycetalesCell Cycle ProteinsMicrotubule-Associated ProteinsSaccharomyces cerevisiae ProteinsLuminescent ProteinsNuclear ProteinsMicroscopy, FluorescenceFluorescence Resonance Energy TransferBinding SitesProtein BindingKineticsMutationModels, BiologicalTime-Lapse ImagingJournal article2018-04-01