Multivalency of NDC80 in the outer kinetochore is essential to track shortening microtubules and generate forces

Presence of multiple copies of the microtubule-binding NDC80 complex is an evolutionary conserved feature of kinetochores, points of attachment of chromosomes to spindle microtubules. This may enable multivalent attachments to microtubules, with implications that remain unexplored. Using recombinant...

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Bibliographic Details
Published in:eLife 2018-04, Vol.7
Main Authors: Volkov, Vladimir A, Huis In 't Veld, Pim J, Dogterom, Marileen, Musacchio, Andrea
Format: Article
Language:English
Subjects:
Binding sites
Biochemistry and Chemical Biology
Cell division
CENP-T
Chromosomal Proteins, Non-Histone - metabolism
Chromosomes
Computer simulation
Cytoskeletal Proteins
Depolymerization
Evolutionary conservation
Experiments
Hec1
HeLa Cells
Humans
kinetochore
Kinetochores
Kinetochores - metabolism
Ligands
Microscopy
microtubule
Microtubule-Associated Proteins - metabolism
Microtubules
Microtubules - physiology
Mitosis
multivalency
Ndc80
Nuclear Proteins - metabolism
Polymerization
Protein Binding
Protein Multimerization
Spindle Apparatus - metabolism
Structural Biology and Molecular Biophysics
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Summary:Presence of multiple copies of the microtubule-binding NDC80 complex is an evolutionary conserved feature of kinetochores, points of attachment of chromosomes to spindle microtubules. This may enable multivalent attachments to microtubules, with implications that remain unexplored. Using recombinant human kinetochore components, we show that while single NDC80 complexes do not track depolymerizing microtubules, reconstituted particles containing the NDC80 receptor CENP-T bound to three or more NDC80 complexes do so effectively, as expected for a kinetochore force coupler. To study multivalency systematically, we engineered modules allowing incremental addition of NDC80 complexes. The modules' residence time on microtubules increased exponentially with the number of NDC80 complexes. Modules with two or more complexes tracked depolymerizing microtubules with increasing efficiencies, and stalled and rescued microtubule depolymerization in a force-dependent manner when conjugated to cargo. Our observations indicate that NDC80, rather than through biased diffusion, tracks depolymerizing microtubules by harnessing force generated during microtubule disassembly.
ISSN:2050-084X
2050-084X
DOI:10.7554/elife.36764