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The polarity and dynamics of microtubule assembly in the budding yeast Saccharomyces cerevisiae

Nature Cell Biology volume 2, pages 36–41 (2000)Cite this article

Abstract

Microtubule assembly in Saccharomyces cerevisiae is initiated from sites within spindle pole bodies (SPBs) in the nuclear envelope. Microtubule plus ends are thought to be organized distal to the SPBs, while minus ends are proximal. Several hypotheses for the function of microtubule motor proteins in force generation and regulation of microtubule assembly propose that assembly and disassembly occur at minus ends as well as at plus ends. Here we analyse microtubule assembly relative to the SPBs in haploid yeast cells expressing green fluorescent protein fused to α-tubulin, a microtubule subunit. Throughout the cell cycle, analysis of fluorescent speckle marks on cytoplasmic astral microtubules reveals that there is no detectable assembly or disassembly at minus ends. After laser-photobleaching, metaphase spindles recover about 63% of the bleached fluorescence, with a half-life of about 1 minute. After anaphase onset, photobleached marks in the interpolar spindle are persistent and do not move relative to the SPBs. In late anaphase, the elongated spindles disassemble at the microtubule plus ends. These results show for astral and anaphase interpolar spindle microtubules, and possibly for metaphase spindle microtubules, that microtubule assembly and disassembly occur at plus, and not minus, ends.

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Figure 1: FSM of an astral microtubule during the G1/S phase of the cell cycle.
Figure 2: FSM of an astral microtubule during anaphase.
Figure 3: Partial fluorescence recovery after laser photobleaching of the metaphase spindle.
Figure 4: Time-lapse observation of an anaphase spindle after laser photobleaching.
Figure 5: FSM of a late-anaphase spindle.
Figure 6: FSM of spindle disassembly in telophase.

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Acknowledgements

P.S.M. thanks members of the Salmon and Bloom laboratories for support and A. Straight for the gift of GFP–Tub1. We also thank Nikon, Hamamatsu Photonics and Universal Imaging for the loan of equipment used for development of our imaging systems. This work was supported by NIH grants GM32238 (to K.S.B.) and GM24364 (to E.D.S.).

Correspondence and requests for materials should be addressed to P.S.M.

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  1. Department of Biology, CB3280, University of North Carolina, Chapel Hill, 27599-3280, North Carolina, USA

    Paul S. Maddox, Kerry S. Bloom & E. D. Salmon

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  1. Paul S. Maddox
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Correspondence to Paul S. Maddox.

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Maddox, P., Bloom, K. & Salmon, E. The polarity and dynamics of microtubule assembly in the budding yeast Saccharomyces cerevisiae. Nat Cell Biol 2, 36–41 (2000). https://doi.org/10.1038/71357

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