EB3-informed dynamics of the microtubule stabilizing cap during stalled growth

  • Microtubule stability is known to be governed by a stabilizing GTP/GDP-Pi cap, but the exact relation between growth velocity, GTP hydrolysis, and catastrophes remains unclear. We investigate the dynamics of the stabilizing cap through in vitro reconstitution of microtubule dynamics in contact with microfabricated barriers, using the plus-end binding protein GFP-EB3 as a marker for the nucleotide state of the tip. The interaction of growing microtubules with steric objects is known to slow down microtubule growth and accelerate catastrophes. We show that the lifetime distributions of stalled microtubules, as well as the corresponding lifetime distributions of freely growing microtubules, can be fully described with a simple phenomenological 1D model based on noisy microtubule growth and a single EB3-dependent hydrolysis rate. This same model is furthermore capable of explaining both the previously reported mild catastrophe dependence on microtubule growth rates and the catastropheMicrotubule stability is known to be governed by a stabilizing GTP/GDP-Pi cap, but the exact relation between growth velocity, GTP hydrolysis, and catastrophes remains unclear. We investigate the dynamics of the stabilizing cap through in vitro reconstitution of microtubule dynamics in contact with microfabricated barriers, using the plus-end binding protein GFP-EB3 as a marker for the nucleotide state of the tip. The interaction of growing microtubules with steric objects is known to slow down microtubule growth and accelerate catastrophes. We show that the lifetime distributions of stalled microtubules, as well as the corresponding lifetime distributions of freely growing microtubules, can be fully described with a simple phenomenological 1D model based on noisy microtubule growth and a single EB3-dependent hydrolysis rate. This same model is furthermore capable of explaining both the previously reported mild catastrophe dependence on microtubule growth rates and the catastrophe statistics during tubulin washout experiments.show moreshow less

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Author:Maurits KokORCiD, Florian HuberORCiD, Svenja-Marei Kalisch, Marileen DogteromORCiD
Qualitätssicherung:peer reviewed
open access:Gold - Erstveröffentlichung mit Lizenzhinweis
Institutes:Zentrum für Digitalisierung und Digitalität (ZDD)
Research fields:Leben und Wohlergehen / Lebewesen
Fachbereich/Einrichtung:Hochschule Düsseldorf / Fachbereich - Medien
Document Type:Article
Year of Completion:2025
Language of Publication:English
Publisher:Elsevier
Parent Title (English):Biophysical Journal
Volume:124
Issue:2
Page Number:18
First Page:227
Last Page:244
URN:urn:nbn:de:hbz:due62-opus-53164
DOI:https://doi.org/10.1016/j.bpj.2024.11.3314
ISSN:1542-0086
GND Keyword:Mikrotubulus
Licence (German):Creative Commons - CC BY-NC-ND - Namensnennung - Nicht kommerziell - Keine Bearbeitungen 4.0 International
Release Date:2025/05/21
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