Measuring NDC80 binding reveals the molecular basis of tension-dependent kinetochore-microtubule attachments.

ABSTRACT: Proper kinetochore-microtubule attachments, mediated by the NDC80 complex, are required for error-free chromosome segregation. Erroneous attachments are corrected by the tension dependence of kinetochore-microtubule interactions. Here, we present a method, based on fluorescence lifetime imaging microscopy and Förster resonance energy transfer, to quantitatively measure the fraction of NDC80 complexes bound to microtubules at individual kinetochores in living human cells. We found that NDC80 binding is modulated in a chromosome autonomous fashion over prometaphase and metaphase, and is predominantly regulated by centromere tension. We show that this tension dependency requires phosphorylation of the N-terminal tail of Hec1, a component of the NDC80 complex, and the proper localization of Au

SUBMITTER: Yoo TY 

PROVIDER: S-EPMC6089600 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature