Abstract
Microtubules (MTs) are important components of the eukaryotic cytoskeleton: they contribute to cell shape and movement, as well as to the motions of organelles including mitotic chromosomes. MTs bind motor enzymes that drive many such movements, but MT dynamics can also contribute to organelle motility1,2,3,4,5,6,7,8. Each MT polymer is a store of chemical energy that can be used to do mechanical work, but how this energy is converted to motility remains unknown. Here we show, by conjugating glass microbeads to tubulin polymers through strong inert linkages, such as biotin–avidin, that depolymerizing MTs exert a brief tug on the beads, as measured with laser tweezers. Analysis of these interactions with a molecular-mechanical model of MT structure and force production9,10 shows that a single depolymerizing MT can generate about ten times the force that is developed by a motor enzyme; thus, this mechanism might be the primary driving force for chromosome motion. Because even the simple coupler used here slows MT disassembly, physiological couplers may modulate MT dynamics in vivo.
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Acknowledgements
We thank A. Hunt and G. J. Bouchard for sharing the plans for their laser tweezers; T. Perkins for advice, the quadrant photo detector design and programs to help with instrument calibration; H. Higuchi for tips on buffers; and members of McIntosh laboratory, and A. I. Vorobjev and G. P. Georgiev for help and support. V. Sarbash, T. Buxkemper and C. Bowen helped with building the trap. This work was supported in part by grants from the NIH to J.R.M., who is a Research Professor of the American Cancer Society.
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Supplementary Notes
This file contains a description of our mathematical model of MT depolymerization, of MT-bead association, and of the laser trap. We calculate the position of the bead and shape of the MT when depolymerization is triggered via two scenarios described in the article. These calculations are compared with experimental results. The last section presents equations that were used to fit the experimental curves (DOC 43 kb)
Supplementary Figures
This file contains Supplementary Figures 1–3 and their legends. The figures display calculated profiles of the bead-MT system and the corresponding graphs. (PDF 499 kb)
Supplementary Video
Each frame shows the result of a calculation of the MT-bead configuration for “depolymerization” of one dimer layer via scenario 1. The bead moves when bending occurs in the bead–associated dimers and the dimers immediately downstream from it. All parameters are as in Supplementary Figure 2, except the attached dimers are # 10–14. The initial configuration is shown in gray. (MOV 132 kb)
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Grishchuk, E., Molodtsov, M., Ataullakhanov, F. et al. Force production by disassembling microtubules. Nature 438, 384–388 (2005). https://doi.org/10.1038/nature04132
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DOI: https://doi.org/10.1038/nature04132
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