The speed of GTP hydrolysis determines GTP cap size and controls microtubule stability

Roostalu, Johanna; Thomas, Claire Le; Cade, Nicholas I.; Kunzelmann, Simone; Taylor, Ian A.; Surrey, Thomas

doi:10.7554/elife.51992

Cited by 93 publications

(128 citation statements)

References 54 publications

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“…A closer examination of MT pauses revealed that these events in control cells were significantly more long-lived: 8.3 ± 5.6 s (mean ± SD) compared to those in KIF21B-KO cells: 4.1 ± 2.2 s (KO #1, mean ± SD) and 4.3 ± 2.4 s (KO #2, mean ± SD) ( Figure 4F , Figure 4—figure supplement 1A ). We note that these pause durations are longer than those observed with EB3-mCherry ( Figure 3I ), because the latter labels growing MTs, and its signal is gradually lost when MT plus ends switch to pausing and lose the GTP cap that EB3 labels ( Roostalu et al, 2020 ). Overall, these data indicate that in T cells, KIF21B acts as a factor that inhibits MT growth and induces pausing and catastrophes.…”

Section: Resultsmentioning

confidence: 85%

Kinesin-4 KIF21B limits microtubule growth to allow rapid centrosome polarization in T cells

Hooikaas

Damstra

Gros

et al. 2020

eLife

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When a T cell and an antigen-presenting cell form an immunological synapse, rapid dynein-driven translocation of the centrosome toward the contact site leads to reorganization of microtubules and associated organelles. Currently, little is known about how the regulation of microtubule dynamics contributes to this process. Here, we show that the knockout of KIF21B, a kinesin-4 linked to autoimmune disorders, causes microtubule overgrowth and perturbs centrosome translocation. KIF21B restricts microtubule length by inducing microtubule pausing typically followed by catastrophe. Catastrophe induction with vinblastine prevented microtubule overgrowth and was sufficient to rescue centrosome polarization in KIF21B-knockout cells. Biophysical simulations showed that a relatively small number of KIF21B molecules can restrict mirotubule length and promote an imbalance of dynein-mediated pulling forces that allows the centrosome to translocate past the nucleus. We conclude that proper control of microtubule length is important for allowing rapid remodeling of the cytoskeleton and efficient T cell polarization.

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Section: Resultsmentioning

confidence: 85%

Kinesin-4 KIF21B limits microtubule growth to allow rapid centrosome polarization in T cells

Hooikaas

Damstra

Gros

et al. 2020

eLife

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“…A cap of GTP-tubulin at the end of a growing microtubule is widely accepted as the determinant of microtubule stability (Mitchison and Kirschner, 1984) (Drechsel and Kirschner, 1994) (Desai and Mitchison, 1997) (Duellberg et al, 2016) (Roostalu et al, 2020). The size of the GTP-cap is defined by the balance between the addition of new GTP-tubulin dimers to the growing microtubule end, and the hydrolysis of GTP to GDP within the microtubule polymer.…”

Section: Discussionmentioning

confidence: 99%

XMAP215 promotes microtubule catastrophe by disrupting the growing microtubule end

Farmer

Arpag

Hall

et al. 2020

Preprint

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The GTP-tubulin cap is widely accepted to protect microtubules against catastrophe. The GTP-cap size is thought to increase with the microtubule growth rate, presumably endowing fast-growing microtubules with enhanced stability. It is unknown what GTP-cap properties permit frequent microtubule catastrophe despite fast growth. Here, we investigate microtubules grown in vitro in the presence and absence of the microtubule polymerase XMAP215. Using EB1 as a GTP-cap marker, we find that GTP-cap size increases regardless of whether growth acceleration is achieved by increasing tubulin concentration or by XMAP215. In spite of the increased mean GTP-cap size, microtubules grown with XMAP215 display increased catastrophe frequency, in contrast to microtubules grown with more tubulin, for which catastrophe is abolished. However, microtubules polymerized with XMAP215 have large fluctuations in growth rate and EB1 intensity; display tapered and curled ends; and undergo catastrophe at faster growth rates and with higher EB1 end-localization. Our results underscore the role of growth irregularities in overall microtubule stability.

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“…Whatever the precise conformational cycle, our results agree best with the view of GMPCPP-tubulin being one of the cap-stabilizing, expanded, and flexible conformations that are unlikely to be preceded by stiffer and compacted ones. Perhaps a more promising approach toward ultimately resolving this issue in future structure determination efforts would be to use knowledge-based point mutations that selectively uncouple the tubulin conformational and GTPase cycles [51,52].…”

Section: Plos Computational Biologymentioning

confidence: 99%

Microtubule instability driven by longitudinal and lateral strain propagation

Igaev

Grubmüller

2020

PLoS Comput Biol

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Tubulin dimers associate longitudinally and laterally to form metastable microtubules (MTs). MT disassembly is preceded by subtle structural changes in tubulin fueled by GTP hydrolysis. These changes render the MT lattice unstable, but it is unclear exactly how they affect lattice energetics and strain. We performed long-time atomistic simulations to interrogate the impacts of GTP hydrolysis on tubulin lattice conformation, lateral inter-dimer interactions, and (non-)local lateral coordination of dimer motions. The simulations suggest that most of the hydrolysis energy is stored in the lattice in the form of longitudinal strain. While not significantly affecting lateral bond stability, the stored elastic energy results in more strongly confined and correlated dynamics of GDP-tubulins, thereby entropically destabilizing the MT lattice.

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The speed of GTP hydrolysis determines GTP cap size and controls microtubule stability

Cited by 93 publications

References 54 publications

Kinesin-4 KIF21B limits microtubule growth to allow rapid centrosome polarization in T cells

Kinesin-4 KIF21B limits microtubule growth to allow rapid centrosome polarization in T cells

XMAP215 promotes microtubule catastrophe by disrupting the growing microtubule end

Microtubule instability driven by longitudinal and lateral strain propagation

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