Antagonism between the dynein and Ndc80 complexes at kinetochores controls the stability of kinetochore-microtubule attachments during mitosis

Mohammed A. Amin, Richard Mckenney, Dileep Varma

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Chromosome alignment and segregation during mitosis require kinetochore-microtubule (kMT) attachments that are mediated by the molecular motor dynein and the kMT-binding complex Ndc80. The Rod-ZW10 -Zwilch (RZZ) complex is central to this coordination as it has an important role in dynein recruitment and has recently been reported to have a key function in the regulation of stable kMT attachments in Caenorhabditis elegans besides its role in activating the spindle assembly checkpoint (SAC). However, the mechanism by which these protein complexes control kMT attachments to drive chromosome motility during early mitosis is still unclear. Here, using in vitro total internal reflection fluorescence microscopy, we observed that higher concentrations of Ndc80 inhibited dynein binding to MTs, providing evidence that Ndc80 and dynein antagonize each other’s function. High-resolution microscopy and siRNA-mediated functional disruption revealed that severe defects in chromosome alignment induced by depletion of dynein or the dynein adapter Spindly are rescued by codepletion of the RZZ component Rod in human cells. Interestingly, rescue of the chromosome alignment defects was independent of Rod function in SAC activation and was accompanied by a remarkable restoration of stable kMT attachments. Furthermore, the chromosome alignment rescue depended on the plus-end- directed motility of centromere protein E (CENP-E) because cells codepleted of CENP-E, Rod, and dynein could not establish stable kMT attachments or align their chromosomes properly. Our findings support the idea that dynein may control the function of the Ndc80 complex in stabilizing kMT attachments directly by interfering with Ndc80 -MT binding or indirectly by controlling the Rod-mediated inhibition of Ndc80.

Original languageEnglish (US)
Pages (from-to)5755-5765
Number of pages11
JournalJournal of Biological Chemistry
Volume293
Issue number16
DOIs
StatePublished - Jan 1 2018

Fingerprint

Dyneins
Kinetochores
Mitosis
Microtubules
Chromosomes
M Phase Cell Cycle Checkpoints
Molecular Motor Proteins
Chromosome Segregation
Defects
Fluorescence microscopy
Caenorhabditis elegans
Fluorescence Microscopy
Small Interfering RNA
Restoration
Microscopy
Microscopic examination
Chemical activation
Cells

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Antagonism between the dynein and Ndc80 complexes at kinetochores controls the stability of kinetochore-microtubule attachments during mitosis. / Amin, Mohammed A.; Mckenney, Richard; Varma, Dileep.

In: Journal of Biological Chemistry, Vol. 293, No. 16, 01.01.2018, p. 5755-5765.

Research output: Contribution to journalArticle

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