Evidence for a HURP/EB free mixed-nucleotide zone in kinetochore-microtubules

Cédric Castrogiovanni, Alessio V. Inchingolo, Jonathan U. Harrison, Damian Dudka, Onur Sen, Nigel J. Burroughs, Andrew D. McAinsh () and Patrick Meraldi ()
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Cédric Castrogiovanni: University of Geneva
Alessio V. Inchingolo: University of Warwick
Jonathan U. Harrison: University of Warwick
Damian Dudka: University of Geneva
Onur Sen: University of Warwick
Nigel J. Burroughs: University of Warwick
Andrew D. McAinsh: University of Warwick
Patrick Meraldi: University of Geneva

Nature Communications, 2022, vol. 13, issue 1, 1-16

Abstract: Abstract Current models infer that the microtubule-based mitotic spindle is built from GDP-tubulin with small GTP caps at microtubule plus-ends, including those that attach to kinetochores, forming the kinetochore-fibres. Here we reveal that kinetochore-fibres additionally contain a dynamic mixed-nucleotide zone that reaches several microns in length. This zone becomes visible in cells expressing fluorescently labelled end-binding proteins, a known marker for GTP-tubulin, and endogenously-labelled HURP - a protein which we show to preferentially bind the GDP microtubule lattice in vitro and in vivo. We find that in mitotic cells HURP accumulates on the kinetochore-proximal region of depolymerising kinetochore-fibres, whilst avoiding recruitment to nascent polymerising K-fibres, giving rise to a growing “HURP-gap”. The absence of end-binding proteins in the HURP-gaps leads us to postulate that they reflect a mixed-nucleotide zone. We generate a minimal quantitative model based on the preferential binding of HURP to GDP-tubulin to show that such a mixed-nucleotide zone is sufficient to recapitulate the observed in vivo dynamics of HURP-gaps.

Date: 2022
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DOI: 10.1038/s41467-022-32421-x

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