Abstract
Microtubules are key players in cellular self-organization, acting as structural scaffolds, cellular highways, force generators and signalling platforms. Microtubules are polar filaments that undergo dynamic instability, i.e. transition between phases of growth and shrinkage. This allows microtubules to explore the inner space of the cell, generate pushing and pulling forces and remodel themselves into arrays with different geometry and function such as the mitotic spindle. To do this, eukaryotic cells employ an arsenal of regulatory proteins to control microtubule dynamics spatially and temporally. Plants and microorganisms have developed secondary metabolites that perturb microtubule dynamics, many of which are in active use as cancer chemotherapeutics and anti-inflammatory drugs. Here, we summarize the methods used to visualize microtubules and to measure the parameters of dynamic instability to study both microtubule regulatory proteins and the action of small molecules interfering with microtubule assembly and/or disassembly.
Highlights
• The proper regulation of microtubule dynamics is essential for faithful chromosome segregation
useful to measure the direct effect of individual proteins
an open access article published by Portland Press Limited on behalf
Summary
We expect that our knowledge in microtubule regulation will increase dramatically over the decade due to several recent breakthroughs: (1) Advances in live cell imaging allow 3D timelapse imaging with unprecedented temporal resolution, (2) the cryoEM revolution enables uncovering the structural changes in tubulin following nucleotide hydrolysis and the interactions of microtubule regulators with tubulin and (3) the purification of recombinant human tubulin enables studying point mutations, modifications and isoforms. Establishing robust assays to measure microtubule dynamics both in vitro and in living cells will be important to push the boundary of our understanding. It is increasingly apparent that different cell types express different compositions of tubulin isoforms, and regulate microtubule dynamics locally and temporally; microtubule dynamics is context dependent. To truly understand how simple tubulin dimers can build the beautiful and complex microtubule network, we must gain context-specific understanding of how tubulin isoforms, tubulin modifications and MAPs guide the fascinating phenomenon of dynamic instability
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