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A new function for the γ -tubulin ring complex as a microtubule minus-end cap

Nature Cell Biology volume 2, pages 358–364 (2000)Cite this article

Abstract

Microtubule nucleation from centrosomes involves a lockwasher-shaped protein complex containing γ-tubulin, named the γ-tubulin ring complex (γTuRC). Here we investigate the mechanism by which the γTuRC nucleates microtubules, using a direct labelling method to visualize the behaviour of individual γTuRCs. A fluorescently-labelled version of the γTuRC binds to the minus ends of microtubules nucleated in vitro. Both γTuRC-mediated nucleation and binding of the γTuRC to preformed microtubules block further minus-end growth and prevent microtubule depolymerization. The γTuRC therefore acts as a minus-end-capping protein, as confirmed by electron-microscopic examination of gold-labelled γTuRCs. These data support a nucleation model for γTuRC function that involves capping of microtubules.

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Figure 1: Covalent attachment of the fluorophore, OGM or biotin to the γTuRC labels all subunits and does not affect in vitro microtubule-nucleating activity.
Figure 2: Microtubule nucleation in the presence of OGM-labelled γTuRCs.
Figure 3: γTuRC binding correlates with microtubule minus-end capping.
Figure 4: Binding of γTuRCs to preformed microtubules prevents minus-end growth.
Figure 5: γTuRC binding prevents minus-end shrinkage.
Figure 6: Electron-microscopic analysis of gold-labelled γTuRCs.
Figure 7: Models for nucleation-independent capping activity of the γTuRC.

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Acknowledgements

We thank J. Heymann for encouragement to try the direct labelling method. C.W. gratefully acknowledges K. Oegema’s help with ‘microtubulology’, and M. Sepanski’s expert technical assistance with the electron microscope. We thank J. Yanowitz, J. Gall, R. Gunawardane and S. Lizarraga for critical reading of the manuscript and A. Fire for help with statistical analysis. This work was supported by a postdoctoral fellowship from the American Cancer Society to C.W., and by a Pew Scholar’s Award and NIH Grant GM56312-01 to Y.Z.

Correspondence and requests for materials should be addressed to C.W.

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  1. Department of Embryology, Carnegie Institution of Washington, 115 West University Parkway, Baltimore, Maryland, 21210, USA

    Christiane Wiese & Yixian Zheng

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Correspondence to Christiane Wiese.

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Wiese, C., Zheng, Y. A new function for the γ -tubulin ring complex as a microtubule minus-end cap. Nat Cell Biol 2, 358–364 (2000). https://doi.org/10.1038/35014051

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