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Willin and Par3 cooperatively regulate epithelial apical constriction through aPKC-mediated ROCK phosphorylation

Nature Cell Biology volume 13, pages 860–866 (2011)Cite this article

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Abstract

Apical-domain constriction is important for regulating epithelial morphogenesis. Epithelial cells are connected by apical junctional complexes (AJCs) that are lined with circumferential actomyosin cables. The contractility of these cables is regulated by Rho-associated kinases (ROCKs). Here, we report that Willin (a FERM-domain protein) and Par3 (a polarity-regulating protein) cooperatively regulate ROCK-dependent apical constriction. We found that Willin recruits aPKC and Par6 to the AJCs, independently of Par3. Simultaneous depletion of Willin and Par3 completely removed aPKC and Par6 from the AJCs and induced apical constriction. Induced constriction was through upregulation of the level of AJC-associated ROCKs, which was due to loss of aPKC. Our results indicate that aPKC phosphorylates ROCK and suppresses its junctional localization, thereby allowing cells to retain normally shaped apical domains. Thus, we have uncovered a Willin/Par3–aPKC–ROCK pathway that controls epithelial apical morphology.

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Figure 1: Willin recruits aPKC to AJCs in MDCK cells.
Figure 2: Willin and Par3 cooperatively regulate aPKC localization and apical constriction in EpH4 cells.
Figure 3: Interaction between aPKC and Par3/Willin is necessary to regulate apical actomyosin activity.
Figure 4: aPKC phosphorylates ROCK1.
Figure 5: ROCK1 phosphorylation regulates its distribution.

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Acknowledgements

We are grateful to K. Shinmyozu for LC-MS/MS analysis. We thank S. Ohno and S. Yonemura for reagents; M. Nomura, H. Saitou and Y. Inoue for technical support; and T. Otani for comments on the research. This work was supported by the programme Grants-in-Aid for Specially Promoted Research of the Ministry of Education, Science, Sports, and Culture of Japan to M.T. T.I. is a recipient of the RIKEN Junior Research Associate fellowship.

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Authors and Affiliations

  1. RIKEN Center for Developmental Biology, Chuo-ku, Kobe 650-0047, Japan

    Takashi Ishiuchi & Masatoshi Takeichi

  2. Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan

    Takashi Ishiuchi

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  1. Takashi Ishiuchi
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T.I. designed and conducted experiments. T.I. and M.T. wrote the paper.

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Correspondence to Masatoshi Takeichi.

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Ishiuchi, T., Takeichi, M. Willin and Par3 cooperatively regulate epithelial apical constriction through aPKC-mediated ROCK phosphorylation. Nat Cell Biol 13, 860–866 (2011). https://doi.org/10.1038/ncb2274

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