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An essential role of the aPKC–Aurora A–NDEL1 pathway in neurite elongation by modulation of microtubule dynamics

Nature Cell Biology volume 11, pages 1057–1068 (2009)Cite this article

Abstract

Orchestrated remodelling of the cytoskeketon is prominent during neurite extension. In contrast with the extensive characterization of actin filament regulation, little is known about the dynamics of microtubules during neurite extension. Here we identify an atypical protein kinase C (aPKC)–Aurora A–NDEL1 pathway that is crucial for the regulation of microtubule organization during neurite extension. aPKC phosphorylates Aurora A at Thr 287 (T287), which augments interaction with TPX2 and facilitates activation of Aurora A at the neurite hillock, followed by phosphorylation of NDEL1 at S251 and recruitment. Suppression of aPKC, Aurora A or TPX2, or disruption of Ndel1, results in severe impairment of neurite extension. Analysis of microtubule dynamics with a microtubule plus-end marker revealed that suppression of the aPKC–Aurora A–NDEL1 pathway resulted in a significant decrease in the frequency of microtubule emanation from the microtubule organizing centre (MTOC), suggesting that Aurora A acts downstream of aPKC. These findings demonstrate a surprising role of aPKC–Aurora A–NDEL1 pathway in microtubule remodelling during neurite extension.

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Figure 1: Aurora A is expressed and phosphorylated in DRG neurons.
Figure 2: PKC-ζ is colocalized with activated Aurora A and phosphorylates Aurora A.
Figure 3: Characterization of phosphorylation of Aurora A on T287 by PKC-ζ using a specific monoclonal antibody against phosphorylated Thr 287.
Figure 4: Phosphorylation of Aurora A on T287 augments interaction with TPX2 and facilitates activation of Aurora A.
Figure 5: Determination of the order of action of aPKC, Aurora A, TPX2 and NDEL1.
Figure 6: An essential role of the PKC-ζ–Aurora A–NDEL1 pathway on neurite elongation.
Figure 7: The PKC-ζ–Aurora A–NDEL1 pathway regulates microtubule projection from the MTOC.
Figure 8: Expression of phosphorylation mimetic Aurora A mutants and neurite extension under suppression of aPKC, depletion of Aurora A, depletion of TPX2 and disruption of Ndel1.

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Acknowledgements

We thank Yukimi Kira, Yoriko Yabunaka, Gaku Kuwabara, Toshiyuki Kawashima and Takako Takitho for technical support, and Hiromichi Nishimura and Keiko Fujimoto for mouse breeding. This work was also supported by the Osaka Community Foundation to D.M. This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan to S.H. This work was also supported by the Cell Science Research Foundation, the Japan Spina Bifida and Hydrocephalus Research Foundation, the Takeda Science Foundation and the Hoh-ansha Foundation to S.H, and National Institutes of Health grants NS41030 and HD47380 to A.W.-B.

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Authors and Affiliations

  1. Department of Genetic Disease Research, Osaka City University Graduate School of Medicine, Asahi-machi 1-4-3, Abeno, 545-8585, Osaka, Japan

    Daisuke Mori, Masami Yamada & Shinji Hirotsune

  2. Research Group for Cytoskeleton and Cell Motility, KAN Research Institute, Inc. 3F, Kobe MI R&D Center, 6-7-3 Minatojima-minamimachi, Chuo-ku, 650-0047, Kobe, Japan

    Yuko Mimori-Kiyosue

  3. Laboratory of Molecular Pharmacology, Biosignal Research Center, Kobe University, 657–8501, Kobe, Japan

    Yasuhito Shirai

  4. Department of Molecular Biology, Yokohama City University Graduate School of Medical Science, 3-9 Fukuura, Kanazawa-ku, Yokohama, 236-0004, Kanagawa, Japan

    Atsushi Suzuki & Shigeo Ohno

  5. Division of Gene Regulation, Institute for Advanced Medical Research, Keio University School of Medicine, Shinanomach 35, Shinjuku, Tokyo, Japan

    Hideaki Saya

  6. Department of Pediatrics and Institute for Human Genetics, UCSF School of Medicine, San Francisco, 94143, California, USA

    Anthony Wynshaw-Boris

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Contributions

D.M., M.Y., Y.S. and A.S. performed the experimental work, Y.M.-K., S.O. and H.S. conducted data analysis, and A.W.-B. and S.H. performed project planning and wrote the manuscript.

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Correspondence to Shinji Hirotsune.

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Mori, D., Yamada, M., Mimori-Kiyosue, Y. et al. An essential role of the aPKC–Aurora A–NDEL1 pathway in neurite elongation by modulation of microtubule dynamics. Nat Cell Biol 11, 1057–1068 (2009). https://doi.org/10.1038/ncb1919

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