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  • Original Paper
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Different behavior of l-Afadin and Neurabin-II during the formation and destruction of cell – cell adherens junction

Oncogene volume 18, pages 1609–1617 (1999)Cite this article

Abstract

We have recently isolated two novel actin filament-binding proteins, l-afadin and neurabin-II and shown that they are localized at cell – cell adherens junction (AJ) in epithelial cells. We found here that l-afadin, neurabin-II, ZO-1, and E-cadherin showed similar and different behavior during the formation and destruction of cell – cell AJ in MDCK cells. In MDCK cells, the accumulation of both l-afadin and E-cadherin, but not that of ZO-1, changed in parallel depending on Rac small G protein activity. Dissociation of MDCK cells by culturing the cells at 2 μM Ca2+ caused rapid endocytosis of E-cadherin, but not that of l-afadin or ZO-1. Addition of phorbol 12-myristate 13-acetate to these dissociated cells formed a tight junction-like structure where ZO-1 and l-afadin, but not neurabin-II or E-cadherin, accumulated. We furthermore found that, in non-epithelial EL cells, which expressed E-cadherin and attached to each other, l-afadin, neurabin-II, ZO-1 and E-cadherin were all localized at AJ. In cadherin-deficient L cells, l-afadin was mainly localized at cell – cell contact sites, but ZO-1 was mainly localized at the tip area of cell processes. Neurabin-II did not accumulate at the plasma membrane area. Neither l-afadin nor neurabin-II significantly interacted with α-, β-catenin, E-cadherin, ZO-1 or occludin.

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Acknowledgements

We thank Drs Sh Tsukita, A Nagafuchi and M Itoh (Kyoto University, Kyoto, Japan) for providing us with the anti-ZO-1 Ab, EL cells, L cells and the GST-fusion vectors, containing α- and β-catenins, the cytoplasmic region of E-cadherin, the N-terminal and C-terminal fragments of ZO-1 and the cytoplasmic region of occludin. We also thank Dr W Birchmeier (Max-Delbruck-Center for Molecular Medicine, Berlin, Germany) for providing us with MDCK cells. The work performed at Osaka University Medical School was supported by grants-in-aid for Scientific Research and for Cancer Research from the Ministry of Education, Science, Sports, and Culture, Japan (1998), by grants-in-aid for Abnormalities in Hormone Receptor Mechanisms and for Aging and Health from the Ministry of Health and Welfare, Japan (1998), and by grants from the Human Frontier Science Program (1998).

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Authors and Affiliations

  1. Department of Molecular Biology and Biochemistry, Osaka University Medical School, 565 – 0871, Suite, Japan

    Toshiaki Sakisaka, Kenji Takaishi & Yoshimi Takai

  2. Takai Biotimer Project, ERATO, Japan Science and Technology Corporation, c/o JCR Pharmaceuticals Co., Ltd., 2-2-10 Murotani, Nishi-ku, Kobe, 651-2241, Japan

    Hiroyuki Nakanishi, Kenichi Takahashi, Kenji Mandai, Masako Miyahara & Ayako Satoh

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  1. Toshiaki Sakisaka
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Sakisaka, T., Nakanishi, H., Takahashi, K. et al. Different behavior of l-Afadin and Neurabin-II during the formation and destruction of cell – cell adherens junction. Oncogene 18, 1609–1617 (1999). https://doi.org/10.1038/sj.onc.1202451

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