EB3-informed dynamics of the microtubule stabilizing cap during stalled growth
Maurits Kok (TU Delft - R&D / Innovation, Kavli institute of nanoscience Delft)
Florian Huber (University of Applied Sciences, Düsseldorf, TU Delft - Applied Sciences, Kavli institute of nanoscience Delft, The Netherlands eScience Center)
Svenja Marei Kalisch (Kavli institute of nanoscience Delft)
Marileen Dogterom (TU Delft - Applied Sciences, Kavli institute of nanoscience Delft)
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Abstract
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 catastrophe statistics during tubulin washout experiments.
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