Termination of Protofilament Elongation by Eribulin Induces Lattice Defects that Promote Microtubule Catastrophes

Harinath Doodhi, Andrea E. Prota, Ruddi Rodríguez-García, Hui Xiao, Daniel W. Custar, Katja Bargsten, Eugene A. Katrukha, Manuel Hilbert, Shasha Hua, Kai Jiang, Ilya Grigoriev, Chia Ping H. Yang, David Cox, Susan Band Horwitz, Lukas C. Kapitein, Anna Akhmanova, Michel O. Steinmetz

Research output: Contribution to journalArticle

42 Scopus citations

Abstract

Microtubules are dynamic polymers built of tubulin dimers that attach in a head-to-tail fashion to form protofilaments, which further associate laterally to form a tube. Asynchronous elongation of individual protofilaments can potentially lead to an altered microtubule-end structure that promotes sudden depolymerization, termed catastrophe [1–4]. However, how the dynamics of individual protofilaments relates to overall growth persistence has remained unclear. Here, we used the microtubule targeting anti-cancer drug Eribulin [5–7] to explore the consequences of stalled protofilament elongation on microtubule growth. Using X-ray crystallography, we first revealed that Eribulin binds to a site on β-tubulin that is required for protofilament plus-end elongation. Based on the structural information, we engineered a fluorescent Eribulin molecule. We demonstrate that single Eribulin molecules specifically interact with microtubule plus ends and are sufficient to either trigger a catastrophe or induce slow and erratic microtubule growth in the presence of EB3. Interestingly, we found that Eribulin increases the frequency of EB3 comet “splitting,” transient events where a slow and erratically progressing comet is followed by a faster comet. This observation possibly reflects the “healing” of a microtubule lattice. Because EB3 comet splitting was also observed in control microtubules in the absence of any drugs, we propose that Eribulin amplifies a natural pathway toward catastrophe by promoting the arrest of protofilament elongation. Doodhi et al. show that Eribulin binds to a site on β-tubulin, which is exposed at the plus ends of microtubules. Binding of single Eribulin molecules induces erratic microtubule growth, catastrophes, and splitting of EB3 comets. The authors propose that Eribulin amplifies a natural catastrophe pathway by inhibiting protofilament elongation.

Original languageEnglish (US)
Pages (from-to)1713-1721
Number of pages9
JournalCurrent Biology
Volume26
Issue number13
DOIs
StatePublished - Jul 11 2016

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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    Doodhi, H., Prota, A. E., Rodríguez-García, R., Xiao, H., Custar, D. W., Bargsten, K., Katrukha, E. A., Hilbert, M., Hua, S., Jiang, K., Grigoriev, I., Yang, C. P. H., Cox, D., Horwitz, S. B., Kapitein, L. C., Akhmanova, A., & Steinmetz, M. O. (2016). Termination of Protofilament Elongation by Eribulin Induces Lattice Defects that Promote Microtubule Catastrophes. Current Biology, 26(13), 1713-1721. https://doi.org/10.1016/j.cub.2016.04.053