Article | Published:

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

Nature Cell Biology volume 11, pages 10571068 (2009) | Download Citation

Subjects

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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    & Cytoskeletal events in growth cone steering. Curr. Opin. Neurobiol. 4, 43–48 (1994).

  2. 2.

    & Making the connection: cytoskeletal rearrangements during growth cone guidance. Cell 83, 171–176 (1995).

  3. 3.

    & Regulation of growth cone actin filaments by guidance cues. J. Neurobiol. 58, 92–102 (2004).

  4. 4.

    et al. Filopodia are required for cortical neurite initiation. Nature Cell Biol. 9, 1347–1359 (2007).

  5. 5.

    et al. Ena/VASP is required for neuritogenesis in the developing cortex. Neuron 56, 441–455 (2007).

  6. 6.

    & Nerve growth cone lamellipodia contain two populations of actin filaments that differ in organization and polarity. J. Cell Biol. 119, 1219–1243 (1992).

  7. 7.

    et al. Mechanism of filopodia initiation by reorganization of a dendritic network. J. Cell Biol. 160, 409–421 (2003).

  8. 8.

    & Regulated actin cytoskeleton assembly at filopodium tips controls their extension and retraction. J. Cell Biol. 146, 1097–1106 (1999).

  9. 9.

    Rho GTPases and actin dynamics in membrane protrusions and vesicle trafficking. Trends Cell Biol. 16, 522–529 (2006).

  10. 10.

    & Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia. Cell 81, 53–62 (1995).

  11. 11.

    & Cytoskeletal dynamics and transport in growth cone motility and axon guidance. Neuron 40, 209–227 (2003).

  12. 12.

    Evolutionary conservation of microtubule-capture mechanisms. Nature Rev. Mol. Cell Biol. 3, 296–304 (2002).

  13. 13.

    et al. Rac1 and Cdc42 capture microtubules through IQGAP1 and CLIP-170. Cell 109, 873–885 (2002).

  14. 14.

    et al. Nanometer targeting of microtubules to focal adhesions. J. Cell Biol. 161, 853–859 (2003).

  15. 15.

    et al. CRMP-2 binds to tubulin heterodimers to promote microtubule assembly. Nature Cell Biol. 4, 583–591 (2002).

  16. 16.

    & Dynamics of axonal microtubules regulate the topology of new membrane insertion into the growing neurites. J. Cell Biol. 143, 1077–1086 (1998).

  17. 17.

    The neurogenetics of lissencephaly. Neurol. Clin. 7, 89–105 (1989).

  18. 18.

    , , & Lissencephaly. A human brain malformation associated with deletion of the LIS1 gene located at chromosome 17p13. J. Am. Med. Assoc. 270, 2838–2842 (1993).

  19. 19.

    et al. Isolation of a Miller–Dieker lissencephaly gene containing G protein beta-subunit-like repeats. Nature 364, 717–721 (1993).

  20. 20.

    Lissencephaly and LIS1: insights into the molecular mechanisms of neuronal migration and development. Clin. Genet. 72, 296–304 (2007).

  21. 21.

    , & Life is a journey: a genetic look at neocortical development. Nature Rev. Genet. 3, 342–355 (2002).

  22. 22.

    et al. NUDEL is a novel Cdk5 substrate that associates with LIS1 and cytoplasmic dynein. Neuron 28, 697–711 (2000).

  23. 23.

    et al. A LIS1/NUDEL/cytoplasmic dynein heavy chain complex in the developing and adult nervous system. Neuron 28, 681–696 (2000).

  24. 24.

    et al. NDEL1 phosphorylation by Aurora-A kinase is essential for centrosomal maturation, separation, and TACC3 recruitment. Mol. Cell. Biol. 27, 352–367 (2007).

  25. 25.

    Nerve growth factors (NGF, BDNF) enhance axonal regeneration but are not required for survival of adult sensory neurons. J. Neurosci. 8, 2394–2405 (1988).

  26. 26.

    , , , & The mitotic serine/threonine kinase Aurora2/AIK is regulated by phosphorylation and degradation. Oncogene 19, 4906–4916 (2000).

  27. 27.

    Eph receptor signalling casts a wide net on cell behaviour. Nature Rev. Mol. Cell Biol. 6, 462–475 (2005).

  28. 28.

    & New exchanges in eph-dependent growth cone dynamics. Neuron 46, 161–163 (2005).

  29. 29.

    The molecular mystery of neuronal migration: FAK and Cdk5. Trends Cell Biol. 14, 1–5 (2004).

  30. 30.

    et al. Dishevelled promotes axon differentiation by regulating atypical protein kinase C. Nature Cell Biol. 9, 743–754 (2007).

  31. 31.

    et al. Microtubule affinity-regulating kinase 2 functions downstream of the PAR-3/PAR-6/atypical PKC complex in regulating hippocampal neuronal polarity. Proc. Natl Acad. Sci. USA 103, 8534–8539 (2006).

  32. 32.

    et al. Activation of protein kinase C zeta by peroxynitrite regulates LKB1-dependent AMP-activated protein kinase in cultured endothelial cells. J. Biol. Chem. 281, 6366–6375 (2006).

  33. 33.

    , , & Inhibition of Aurora A in response to DNA damage. Oncogene 25, 338–348 (2006).

  34. 34.

    et al. Human TPX2 is required for targeting Aurora-A kinase to the spindle. J. Cell Biol. 158, 617–623 (2002).

  35. 35.

    , , & Structural basis of Aurora-A activation by TPX2 at the mitotic spindle. Mol. Cell 12, 851–862 (2003).

  36. 36.

    et al. An essential function of the C. elegans ortholog of TPX2 is to localize activated Aurora A kinase to mitotic spindles. Dev. Cell 9, 237–248 (2005).

  37. 37.

    , , & TPX2, A novel Xenopus MAP involved in spindle pole organization. J. Cell Biol. 149, 1405–1418 (2000).

  38. 38.

    et al. EGF or PDGF receptors activate atypical PKCλ through phosphatidylinositol 3-kinase. EMBO J. 15, 788–798 (1996).

  39. 39.

    et al. Requirement of atypical protein kinase Cλ for insulin stimulation of glucose uptake but not for Akt activation in 3T3-L1 adipocytes. Mol. Cell. Biol. 18, 6971–6982 (1998).

  40. 40.

    , & Linking cell cycle to asymmetric division: Aurora-A phosphorylates the Par complex to regulate Numb localization. Cell 135, 161–173 (2008).

  41. 41.

    , & The dynamic behavior of the APC-binding protein EB1 on the distal ends of microtubules. Curr. Biol. 10, 865–868 (2000).

  42. 42.

    et al. Centrosome localization determines neuronal polarity. Nature 436, 704–708 (2005).

  43. 43.

    et al. LIS1 and NDEL1 coordinate the plus-end-directed transport of cytoplasmic dynein. EMBO J. 27, 2471–2483 (2008).

  44. 44.

    & Asymmetric cell division in the Drosophila nervous system. Nature Rev. Neurosci. 2, 772–779 (2001).

  45. 45.

    Intercellular junctions and cellular polarity: the PAR-aPKC complex, a conserved core cassette playing fundamental roles in cell polarity. Curr. Opin. Cell Biol. 13, 641–648 (2001).

  46. 46.

    , & Hippocampal neuronal polarity specified by spatially localized mPar3/mPar6 and PI 3-kinase activity. Cell 112, 63–75 (2003).

  47. 47.

    et al. Drosophila Aurora-A kinase inhibits neuroblast self-renewal by regulating aPKC/Numb cortical polarity and spindle orientation. Genes Dev. 20, 3464–3474 (2006).

  48. 48.

    et al. Aurora-A acts as a tumor suppressor and regulates self-renewal of Drosophila neuroblasts. Genes Dev. 20, 3453–3463 (2006).

  49. 49.

    , , , & HEF1-dependent Aurora A activation induces disassembly of the primary cilium. Cell 129, 1351–1363 (2007).

  50. 50.

    et al. A centrosomal Cdc20-APC pathway controls dendrite morphogenesis in postmitotic neurons. Cell 136, 322–336 (2009).

  51. 51.

    , , & Optimized survival of hippocampal neurons in B27-supplemented Neurobasal, a new serum-free medium combination. J. Neurosci. Res. 35, 567–576 (1993).

Download references

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.

Author information

Affiliations

  1. Department of Genetic Disease Research, Osaka City University Graduate School of Medicine Asahi-machi 1-4-3 Abeno, Osaka 545-8585, 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, Kobe 650-0047, Japan.

    • Yuko Mimori-Kiyosue
  3. Laboratory of Molecular Pharmacology, Biosignal Research Center, Kobe University, Kobe 657–8501, Japan.

    • Yasuhito Shirai
  4. Department of Molecular Biology, Yokohama City University Graduate School of Medical Science, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, 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. UCSF School of Medicine, Department of Pediatrics and Institute for Human Genetics, San Francisco, California 94143, USA.

    • Anthony Wynshaw-Boris

Authors

  1. Search for Daisuke Mori in:

  2. Search for Masami Yamada in:

  3. Search for Yuko Mimori-Kiyosue in:

  4. Search for Yasuhito Shirai in:

  5. Search for Atsushi Suzuki in:

  6. Search for Shigeo Ohno in:

  7. Search for Hideaki Saya in:

  8. Search for Anthony Wynshaw-Boris in:

  9. Search for Shinji Hirotsune in:

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.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Shinji Hirotsune.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Information

Videos

  1. 1.

    Supplementary Information

    Supplementary Movie 1

  2. 2.

    Supplementary Information

    Supplementary Movie 2

  3. 3.

    Supplementary Information

    Supplementary Movie 3

  4. 4.

    Supplementary Information

    Supplementary Movie 4

  5. 5.

    Supplementary Information

    Supplementary Movie 5

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ncb1919

Further reading