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AMPK controls the speed of microtubule polymerization and directional cell migration through CLIP-170 phosphorylation

Nature Cell Biology volume 12, pages 583–590 (2010)Cite this article

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Abstract

AMP-activated protein kinase (AMPK) is an energy-sensing Ser/Thr protein kinase originally shown to be regulated by AMP1. AMPK is activated by various cellular stresses that inhibit ATP production or stimulate ATP consumption2. In addition to its role in metabolism, AMPK has recently been reported to reshape cells by regulating cell polarity and division3,4,5,6. However, the downstream targets of AMPK that participate in these functions have not been fully identified. Here, we show that phosphorylation of the microtubule plus end protein CLIP-170 by AMPK is required for microtubule dynamics and the regulation of directional cell migration. Both inhibition of AMPK and expression of a non-phosphorylatable CLIP-170 mutant resulted in prolonged and enhanced accumulation of CLIP-170 at microtubule tips, and slower tubulin polymerization. Furthermore, inhibition of AMPK impaired microtubule stabilization and perturbed directional cell migration. All of these phenotypes were rescued by expression of a phosphomimetic CLIP-170 mutant. Our results demonstrate, therefore, that AMPK controls basic cellular functions by regulating microtubule dynamics through CLIP-170 phosphorylation.

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Figure 1: In vitro phosphorylation of CLIP-170 Ser311 by AMPK.
Figure 2: CLIP-170 phosphorylated by AMPK localizes to microtubule tips.
Figure 3: AMPK-phosphorylated CLIP-170 regulates microtubule dynamics.
Figure 4: Loss of CLIP-170 phosphorylation increases the size of focal adhesions.
Figure 5: Phosphorylation of CLIP-170 at Ser 311 is essential for cell polarity and directional cell migration.

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Acknowledgements

We thank M. Amano and S. Fukuhara for helpful discussions, and M. Koyama (Olympus Corporation) for technical advice regarding microscopy. This research was supported by: a Grants-in-Aid from the Ministry of Health, Labour and Welfare of Japan; Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan; grants from the Japan Heart Foundation; grants from the Japan Cardiovascular Research Foundation; a grant from the Japan Society for the Promotion of Science; a grant from the Mochida Memorial Foundation for Medical and Pharmaceutical Research; and a Grant-in-Aid from the Japan Medical Association.

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Authors and Affiliations

  1. Division of Cardiovascular Medicine, National Cardiovascular Center Research Institute Suita, Osaka, 565-8565, Japan

    Atsushi Nakano, Kyung-Duk Min, Satoru Yamazaki, Osamu Seguchi, Hiroshi Asanuma, Masanori Asakura & Masafumi Kitakaze

  2. Department of Structural Analysis, National Cardiovascular Center, Research Institute Suita, Osaka, 565-8565, Japan

    Naoki Mochizuki

  3. Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine Suita, Osaka, 565-0871, Japan

    Hisakazu Kato, Kyung-Duk Min, Yoshihiro Asano, Shuichiro Higo, Yasunori Shintani, Tetsuo Minamino & Seiji Takashima

  4. Department of Molecular Cardiology, Osaka University Graduate School of Medicine Suita, Osaka, 565-0871, Japan

    Atsushi Nakano, Hisakazu Kato, Yoshihiro Asano & Seiji Takashima

  5. Institute for Advanced Research, Nagoya University Graduate School of Medicine, Nagoya, 466-8550, Aichi, Japan

    Takashi Watanabe

  6. Department of Cell Pharmacology, Nagoya University Graduate School of Medicine, Nagoya, 466-8550, Aichi, Japan

    Takashi Watanabe & Kozo Kaibuchi

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  1. Atsushi Nakano
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Contributions

A.N. designed and conducted the study, performed most of the experiments, and wrote the manuscript; S.T. designed and conducted the study, performed the biochemical experiments and wrote the manuscript; H.K. carried out immunoblot analysis; K.M. independently counted the number of cells; S.Y. helped to generate the plasmids; Y.A., O.S., S.H., Y.S., H.A., M.A. and T.M. discussed the results and reviewed the manuscript; T.W. and K.K. generated and provided antibodies and Vero cells and reviewed the manuscript; N.M. conducted and supported the biological experiments and wrote the manuscript; M.K. supervised all work.

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Correspondence to Seiji Takashima.

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The authors declare no competing financial interests.

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Nakano, A., Kato, H., Watanabe, T. et al. AMPK controls the speed of microtubule polymerization and directional cell migration through CLIP-170 phosphorylation. Nat Cell Biol 12, 583–590 (2010). https://doi.org/10.1038/ncb2060

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