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EB1 regulates microtubule dynamics and tubulin sheet closure in vitro

Nature Cell Biology volume 10, pages 415–421 (2008)Cite this article

Abstract

End binding 1 (EB1) is a plus-end-tracking protein (+TIP) that localizes to microtubule plus ends where it modulates their dynamics and interactions with intracellular organelles1,2. Although the regulating activity of EB1 on microtubule dynamics has been studied in cells3,4,5,6 and purified systems7,8,9,10,11,12,13, the molecular mechanisms involved in its specific activity are still unclear. Here, we describe how EB1 regulates the dynamics and structure of microtubules assembled from pure tubulin. We found that EB1 stimulates spontaneous nucleation and growth of microtubules, and promotes both catastrophes (transitions from growth to shrinkage) and rescues (reverse events). Electron cryomicroscopy showed that EB1 induces the initial formation of tubulin sheets, which rapidly close into the common 13-protofilament-microtubule architecture. Our results suggest that EB1 favours the lateral association of free tubulin at microtubule-sheet edges, thereby stimulating nucleation, sheet growth and closure. The reduction of sheet length at microtubule growing-ends together with the elimination of stressed microtubule lattices may account for catastrophes. Conversely, occasional binding of EB1 to the microtubule lattice may induce rescues.

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Figure 1: EB1 stimulates catastrophes and rescues.
Figure 2: EB1 stimulates microtubule self-assembly and promotes sheet formation and closure.
Figure 3: EB1 favours unskewed 13-protofilament microtubules while eliminating lattice defects.
Figure 4: Model for EB1 activity on microtubule assembly and dynamics.

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Acknowledgements

We are very grateful to Niels Galjart for providing us with the mouse EB1 construct. We thank Didier Job, Fabrice Senger, Daniel Thomas and Odile Valiron for helpful discussions, and Jean-Paul Rolland for technical assistance with electron microscopy. This work was supported by grants from the Centre National de la Recherche Scientifique (CNRS), the Ministère de l'Enseignement Supérieur et de la Recherche (MESR) and Rennes Métropole. B. V. was supported by a predoctoral fellowship from the MESR and F. M. C. was supported by a postdoctoral fellowship from the CNRS.

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  1. Université de Rennes 1, Interactions Cellulaires et Moléculaires, Centre Nationale de la Recherche Scientifique, Unité Mixte de Recherche 6026, Institut Fédératif de Recherche 140, Génétique Fonctionnelle Agronomie et Santé, Campus de Beaulieu, 263, Avenue du Général Leclerc, Rennes, 35042, France

    Benjamin Vitre, Frédéric M. Coquelle, Claire Heichette, Cyrille Garnier, Denis Chrétien & Isabelle Arnal

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Correspondence to Denis Chrétien or Isabelle Arnal.

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Supplementary Figures S1 and S2, Supplementary Table 1 and Supplementary Methods (PDF 456 kb)

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Vitre, B., Coquelle, F., Heichette, C. et al. EB1 regulates microtubule dynamics and tubulin sheet closure in vitro. Nat Cell Biol 10, 415–421 (2008). https://doi.org/10.1038/ncb1703

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