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Golgi-derived CLASP-dependent microtubules control Golgi organization and polarized trafficking in motile cells

Nature Cell Biology volume 11, pages 1069–1080 (2009)Cite this article

Abstract

Microtubules are indispensable for Golgi complex assembly and maintenance, which are integral parts of cytoplasm organization during interphase in mammalian cells. Here, we show that two discrete microtubule subsets drive two distinct, yet simultaneous, stages of Golgi assembly. In addition to the radial centrosomal microtubule array, which positions the Golgi in the centre of the cell, we have identified a role for microtubules that form at the Golgi membranes in a manner dependent on the microtubule regulators CLASPs. These Golgi-derived microtubules draw Golgi ministacks together in tangential fashion and are crucial for establishing continuity and proper morphology of the Golgi complex. We propose that specialized functions of these two microtubule arrays arise from their specific geometries. Further, we demonstrate that directional post-Golgi trafficking and cell migration depend on Golgi-associated CLASPs, suggesting that correct organization of the Golgi complex by microtubules is essential for cell polarization and motility.

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Figure 1: The two-stage process of Golgi assembly requires CLASPs.
Figure 2: Golgi assembly occurs in 2 stages after mitotic exit.
Figure 3: Golgi ministack clustering by Golgi-derived and centrosomal microtubules.
Figure 4: Golgi assembly depends on directionality of two microtubule subsets and on dynein activity.
Figure 5: CLASPs at the Golgi determine ribbon morphology.
Figure 6: Golgi fragmentation in CLASP-depleted cells results in diminished enzyme mobility within the Golgi complex.
Figure 7: CLASP-dependent microtubules polarize trafficking to the cell front.
Figure 8: CLASP-dependent microtubules regulate directional cell migration.

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Acknowledgements

We thank James Goldenring, Mathew Tyska and Ryoma Ohi for helpful discussions, and Anna Akhmanova, Laura A. Lee and Steven K. Hanks for critical reading of the manuscript. This study was funded by NIH NIGMS grant 1R01GM078373-01 to I.K. and a pilot project to I.K. from NIH NCI GI SPORE grant P50CA095103. A.R.R.M. was supported by Fundação para a Ciência e Tecnologia fellowship SFRH/BD/32976/2006 and by grant PTDC/SAU-OBD/66113/2006 from Fundacao para a Ciencia e a Tecnologia of Portugal. P.M.M. was supported by American Heart Predoctoral grant 09PRE2260729.

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  1. Andrey Efimov

    Present address: Current address: Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, TN 37232, USA.,

Authors and Affiliations

  1. Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, 37232, TN, USA

    Paul M. Miller, Andrew W. Folkmann, Ana R. R. Maia, Nadia Efimova, Andrey Efimov & Irina Kaverina

  2. Institute for Molecular and Cell Biology, University of Porto, 4150-180, Porto, Portugal

    Ana R. R. Maia

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Contributions

P.M.M. performed most experiments and data analysis and co-wrote the manuscript. A.W.F. contributed to experiments and data quantification. A.R.R.M. contributed to mitosis experiments. N.E. contributed to experiments and provided technical assistance. A.E. performed microtubule tip tracking experiments for Fig. 4a, b. I.K. contributed to experiments, directed all of the work and project planning, and co-wrote the manuscript.

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Correspondence to Irina Kaverina.

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Miller, P., Folkmann, A., Maia, A. et al. Golgi-derived CLASP-dependent microtubules control Golgi organization and polarized trafficking in motile cells. Nat Cell Biol 11, 1069–1080 (2009). https://doi.org/10.1038/ncb1920

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