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

• Published in: eLife Vol. 7
• Main Authors: Volkov, Vladimir A, Huis In 't Veld, Pim J, Dogterom, Marileen, Musacchio, Andrea
• Format: Journal Article
• Language: English
• Published: England eLife Sciences Publications Ltd 09.04.2018; eLife Sciences Publications, Ltd
• 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; microtubule; chemical biology; biochemistry; kinetochore; Hec1; structural biology; molecular biophysics; human; CENP-T; multivalency; Ndc80
• ISSN: 2050-084X; 2050-084X
• DOI: 10.7554/elife.36764

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.