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