Brief Report

Structural Transitions at Microtubule Ends Correlate with Their Dynamic Properties in Xenopus Egg Extracts

Correspondence to: Anthony A. Hyman, EMBL, Meyerhofstrasse 1, 69117 Heidelberg, Germany. Tel:49-6221-387-337 Fax:49-6221-387-512 E-mail:hyman{at}embl-heidelberg.de.

Microtubules are dynamically unstable polymers that interconvert stochastically between growing and shrinking states by the addition and loss of subunits from their ends. However, there is little experimental data on the relationship between microtubule end structure and the regulation of dynamic instability. To investigate this relationship, we have modulated dynamic instability in Xenopus egg extracts by adding a catastrophe-promoting factor, Op18/stathmin. Using electron cryomicroscopy, we find that microtubules in cytoplasmic extracts grow by the extension of a two- dimensional sheet of protofilaments, which later closes into a tube. Increasing the catastrophe frequency by the addition of Op18/stathmin decreases both the length and frequency of the occurrence of sheets and increases the number of frayed ends. Interestingly, we also find that more dynamic populations contain more blunt ends, suggesting that these are a metastable intermediate between shrinking and growing microtubules. Our results demonstrate for the first time that microtubule assembly in physiological conditions is a two-dimensional process, and they suggest that the two-dimensional sheets stabilize microtubules against catastrophes. We present a model in which the frequency of catastrophes is directly correlated with the structural state of microtubule ends.

Key Words: microtubules, dynamic instability, Xenopus egg extracts, electron cryomicroscopy, protofilament sheets

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