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CRMP-2 binds to tubulin heterodimers to promote microtubule assembly

Nature Cell Biology volume 4, pages 583–591 (2002)Cite this article

Abstract

Regulated increase in the formation of microtubule arrays is thought to be important for axonal growth. Collapsin response mediator protein-2 (CRMP-2) is a mammalian homologue of UNC-33, mutations in which result in abnormal axon termination. We recently demonstrated that CRMP-2 is critical for axonal differentiation. Here, we identify two activities of CRMP-2: tubulin-heterodimer binding and the promotion of microtubule assembly. CRMP-2 bound tubulin dimers with higher affinity than it bound microtubules. Association of CRMP-2 with microtubules was enhanced by tubulin polymerization in the presence of CRMP-2. The binding property of CRMP-2 with tubulin was apparently distinct from that of Tau, which preferentially bound microtubules. In neurons, overexpression of CRMP-2 promoted axonal growth and branching. A mutant of CRMP-2, lacking the region responsible for microtubule assembly, inhibited axonal growth and branching in a dominant-negative manner. Taken together, our results suggest that CRMP-2 regulates axonal growth and branching as a partner of the tubulin heterodimer, in a different fashion from traditional MAPs.

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Figure 1: Identification of tubulin as a CRMP-2-interacting protein.
Figure 2: CRMP-2 binds directly to tubulin heterodimers.
Figure 3: Dark-field video microscopy of assembling microtubules in tubulin–CRMP-2 preparations.
Figure 4: CRMP-2 associates with tubulin dimers more efficiently than with microtubules.
Figure 5: CRMP-2 induces neurite formation in N1E-115 cells.
Figure 6: CRMP-2 interacts with soluble tubulin heterodimers in cultured hippocampal neurons.
Figure 7: Overexpression of CRMP-2WT and CRMP-2-Δ323–381 in hippocampal neurons.

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Acknowledgements

We thank Y. Gu, Y. Ihara, Y. Kanai, N. Hirokawa, N. J. Cowan, E. Mekada, T. Kato and M. Nakafuku for kind gifts of materials. We also thank M. Amano, M. Fukata, S. Taya, Y. Kawano (Nagoya University) and H. Qadota (Nara Institute of Science and Technology) for helpful discussion and for preparing some materials, and T. Ishii and M. Yoshizaki for secretarial and technical assistance. This research was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan, the Japan Society for the Promotion of Science Research for the Future and the Human Frontier Science Program. F.Y. is a research fellow of the Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, 65 Tsurumai, Showa, 466-8550, Nagoya, Japan

    Yuko Fukata, Toshihide Kimura, Céline Ménager, Takashi Nishimura, Takashi Shiromizu, Hiroyasu Watanabe & Kozo Kaibuchi

  2. Division of Biological Sciences, Graduate School of Science, Nagoya University, Furo, Chikusa, 464-8602, Nagoya, Japan

    Tomohiko J. Itoh & Hirokazu Hotani

  3. Division of Signal Transduction, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, 630-0101, Japan

    Takashi Shiromizu & Naoyuki Inagaki

  4. Central Laboratories for Key Technology, Kirin Brewery Co. Ltd., 1-13-5 Fukuura, Kanazawa, 236-0004, Yokohama, Japan

    Akihiro Iwamatsu

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Correspondence to Kozo Kaibuchi.

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Figure S1 Direct interaction between CRMP-2 and tubulin heterodimers. (PDF 193 kb)

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Fukata, Y., Itoh, T., Kimura, T. et al. CRMP-2 binds to tubulin heterodimers to promote microtubule assembly. Nat Cell Biol 4, 583–591 (2002). https://doi.org/10.1038/ncb825

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