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

36 Citations (Scopus)

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

Fingerprint

eribulin
Crystal defects
Microtubules
microtubules
Elongation
Tubulin
tubulin
Growth
Molecules
Depolymerization
X ray crystallography

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., ... 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

Termination of Protofilament Elongation by Eribulin Induces Lattice Defects that Promote Microtubule Catastrophes. / Doodhi, Harinath; Prota, Andrea E.; Rodríguez-García, Ruddi; Xiao, Hui; Custar, Daniel W.; Bargsten, Katja; Katrukha, Eugene A.; Hilbert, Manuel; Hua, Shasha; Jiang, Kai; Grigoriev, Ilya; Yang, Chia-Ping H.; Cox, David; Band Horwitz, Susan; Kapitein, Lukas C.; Akhmanova, Anna; Steinmetz, Michel O.

In: Current Biology, 11.04.2015.

Research output: Contribution to journalArticle

Doodhi, H, Prota, AE, Rodríguez-García, R, Xiao, H, Custar, DW, Bargsten, K, Katrukha, EA, Hilbert, M, Hua, S, Jiang, K, Grigoriev, I, Yang, C-PH, Cox, D, Band Horwitz, S, Kapitein, LC, Akhmanova, A & Steinmetz, MO 2015, '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
Doodhi, Harinath ; Prota, Andrea E. ; Rodríguez-García, Ruddi ; Xiao, Hui ; Custar, Daniel W. ; Bargsten, Katja ; Katrukha, Eugene A. ; Hilbert, Manuel ; Hua, Shasha ; Jiang, Kai ; Grigoriev, Ilya ; Yang, Chia-Ping H. ; Cox, David ; Band Horwitz, Susan ; Kapitein, Lukas C. ; Akhmanova, Anna ; Steinmetz, Michel O. / Termination of Protofilament Elongation by Eribulin Induces Lattice Defects that Promote Microtubule Catastrophes. In: Current Biology. 2015.
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