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Caenorhabditis elegans EFA-6 limits microtubule growth at the cell cortex

Nature Cell Biology volume 12, pages 1235–1241 (2010)Cite this article

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Abstract

Microtubules are polymers of tubulin heterodimers that exhibit dynamic instability: periods of growth followed by periods of shrinkage1. However, the molecular regulation of dynamic instability remains elusive. Here, we show that EFA-6, a cortically-localized protein2, limits the growth of microtubules near the cell cortex of early embryonic cells from Caenorhabidits elegans, possibly by inducing microtubule catastrophes. Compared with wild type, embryos lacking EFA-6 had abnormally long and dense microtubules at the cell cortex, and growing microtubule plus ends resided at the cortex for up to five-fold longer. Loss of EFA-6 also caused excess centrosome separation and displacement towards the cell cortex early in mitosis, and subsequently a loss of anaphase spindle-pole oscillations and increased rates of spindle elongation. The centrosome separation phenotype was dependent on the motor protein dynein, suggesting a possible link between the modulation of microtubule dynamics at the cortex and dynein-dependent force production. EFA-6 orthologues activate ARF6-type GTPases to regulate vesicle trafficking3. However, we show that only the C. elegans EFA-6 amino-terminus is both necessary and sufficient to limit microtubule growth along the cortex, and that this function is independent of ARF-6.

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Figure 1: Loss of EFA-6 affects early microtubule-dependent processes.
Figure 2: EFA-6 prevents long cortical microtubules in one- and two-cell embryos.
Figure 3: EFA-6 limits microtubule growth at the cortex.
Figure 4: Determination of the EFA-6 microtubule-regulating residues.

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Acknowledgements

We thank M. Price, G. von Dassow, L. Pelletier, and C. Doe for insightful discussions; M. Goulding and E. Munro for technical advice; S. Schneider for advice on taxonomical and sequence alignments; S. Mitani for isolating and providing efa-6(tm3124); V. Davis Haug for the mC-TBA-1 strain; and C. Doe and C. Cabernard for comments on the manuscript. S.M.O was supported by the Leukemia and Lymphoma Society of America, S.M.O. and B.B. are supported by NIH R01GM058017 and R01GMO49869.

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  1. Institute of Molecular Biology, 1229 University of Oregon, Eugene, 97403, Oregon, USA

    Sean M. O'Rourke, Sara N. Christensen & Bruce Bowerman

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  1. Sean M. O'Rourke
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  2. Sara N. Christensen
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  3. Bruce Bowerman
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Contributions

S.M.O. and B.B. designed the experiments and wrote the paper. S.M.O. conceived the project, performed the experiments and analysed the data. Microscopy and strain generation were also contributed by S.N.C.

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Correspondence to Sean M. O'Rourke or Bruce Bowerman.

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O'Rourke, S., Christensen, S. & Bowerman, B. Caenorhabditis elegans EFA-6 limits microtubule growth at the cell cortex. Nat Cell Biol 12, 1235–1241 (2010). https://doi.org/10.1038/ncb2128

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