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

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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)
JournalCurrent Biology
DOIs
StateAccepted/In press - Apr 11 2015

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ASJC Scopus subject areas

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

Cite this

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., Band Horwitz, S., Kapitein, L. C., Akhmanova, A., & Steinmetz, M. O. (Accepted/In press). Termination of Protofilament Elongation by Eribulin Induces Lattice Defects that Promote Microtubule Catastrophes. Current Biology. https://doi.org/10.1016/j.cub.2016.04.053