TY - JOUR
T1 - Termination of Protofilament Elongation by Eribulin Induces Lattice Defects that Promote Microtubule Catastrophes
AU - Doodhi, Harinath
AU - Prota, Andrea E.
AU - Rodríguez-García, Ruddi
AU - Xiao, Hui
AU - Custar, Daniel W.
AU - Bargsten, Katja
AU - Katrukha, Eugene A.
AU - Hilbert, Manuel
AU - Hua, Shasha
AU - Jiang, Kai
AU - Grigoriev, Ilya
AU - Yang, Chia Ping H.
AU - Cox, David
AU - Horwitz, Susan Band
AU - Kapitein, Lukas C.
AU - Akhmanova, Anna
AU - Steinmetz, Michel O.
N1 - Publisher Copyright:
© 2016 Elsevier Ltd
PY - 2016/7/11
Y1 - 2016/7/11
N2 - 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.
AB - 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.
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U2 - 10.1016/j.cub.2016.04.053
DO - 10.1016/j.cub.2016.04.053
M3 - Article
C2 - 27321995
AN - SCOPUS:85008240103
SN - 0960-9822
VL - 26
SP - 1713
EP - 1721
JO - Current Biology
JF - Current Biology
IS - 13
ER -