Microtubule architecture in vitro and in cells revealed by cryo-electron tomography

Atherton, J. and Stouffer, M. and Francis, F. and Moores, Carolyn A. (2018) Microtubule architecture in vitro and in cells revealed by cryo-electron tomography. Acta Crystallographica Section D: Structural Biology D74 , pp. 572-584. ISSN 2059-7983.

Abstract

The microtubule cytoskeleton is involved in many vital cellular processes. Microtubules act as tracks for molecular motors, and their polymerisation and depolymerisation can be harnessed to generate force. The structures of microtubules provide key information about the mechanisms by which their cellular roles are accomplished and the physiological context in which these roles are performed. Cryo-electron microscopy allows visualisation of in vitro polymerised microtubules and has provided important insights into their overall morphology and the influence of a range factors on their structure and dynamics. Cryo-electron tomography can be used to determine the unique 3D structure of individual microtubules and their ends. Here we revisit our previous cryo-electron tomography study of in vitro polymerised GMPCPP-stabilised microtubules, compare our findings with new tomograms of dynamic in vitro and cellular microtubules and highlight the information that can be extracted from such data. Our analysis shows the surprising structural heterogeneity of in vitro polymerised microtubules. Lattice defects can be observed both in vitro and in cells. The shared ultrastructural properties in these different populations emphasise the relevance of 3D structures of in vitro microtubules for understanding microtubule cellular functions.

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