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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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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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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DOI: https://doi.org/10.1038/ncb825
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