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© The Rockefeller University Press, 0021-9525/1998//847 $5.00
The Journal of Cell Biology, Volume 142, Number 3, , 1998 847-857

Regular Article

{alpha}-Catenin-Vinculin Interaction Functions to Organize the Apical Junctional Complex in Epithelial Cells



Mitsuko Watabe-Uchida*, Naoshige Uchida*, Yuzo Imamura{ddagger}, Akira Nagafuchi{ddagger}, Kazushi Fujimoto§, Tadashi Uemura*, Stefan Vermeulen||, Frans van Roy, Eileen D. Adamson**, and Masatoshi Takeichi*

* Department of Biophysics, Faculty of Science, Kyoto University, Kitashirakawa, Sakyo-ku, Kyoto 606-8502; {ddagger} Department of Cell Biology and § Department of Anatomy, Faculty of Medicine, Kyoto University, Yoshida, Sakyo-ku, Kyoto 606-8507, Japan; || University Hospital of Ghent and Department of Molecular Biology, VIB-University of Ghent, B-9000 Ghent, Belgium; and ** The Burnham Institute, La Jolla, California

{alpha}E-catenin, a cadherin-associated protein, is required for tight junction (TJ) organization, but its role is poorly understood. We transfected an {alpha}E-catenin–deficient colon carcinoma line with a series of {alpha}E-catenin mutant constructs. The results showed that the amino acid 326–509 domain of this catenin was required to organize TJs, and its COOH-terminal domain was not essential for this process. The 326–509 internal domain was found to bind vinculin. When an NH2-terminal {alpha}E-catenin fragment, which is by itself unable to organize the TJ, was fused with the vinculin tail, this chimeric molecule could induce TJ assembly in the {alpha}E-catenin–deficient cells. In vinculin-null F9 cells, their apical junctional organization was impaired, and this phenotype was rescued by reexpression of vinculin. These results indicate that the {alpha}E-catenin-vinculin interaction plays a role in the assembly of the apical junctional complex in epithelia.

Key Words: cadherin • catenin • tight junction • vinculin • zonula adherens

Abbreviations used in this paper: GST, glutathione S-transferase; MBP, maltose-binding proteins; pAb, polyclonal antibody; TJ, tight junction; ZA, zonula adherens.

Address all correspondence to Masatoshi Takeichi, Department of Biophysics, Faculty of Science, Kyoto University, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan. Tel.: 81-75-753-4196; Fax: 81-75-753-4197; E-mail: takeichi{at}take.biophys.kyoto-u.ac.jp


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