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Published online 4 September 2000. doi:10.1083/jcb.150.5.1161
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© The Rockefeller University Press, 0021-9525/2000//1161 $5.00
The Journal of Cell Biology, Volume 150, Number 5, , 2000 1161-1176

Original Article

Two Cell Adhesion Molecules, Nectin and Cadherin, Interact through Their Cytoplasmic Domain–Associated Proteins



Kouichi Tachibanaa, Hiroyuki Nakanishia, Kenji Mandaia, Kumi Ozakia, Wataru Ikedaa, Yasunori Yamamotoa, Akira Nagafuchib, Shoichiro Tsukitab, and Yoshimi Takaia

a Department of Molecular Biology and Biochemistry, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Japan
b Department of Cell Biology, Faculty of Medicine, Kyoto University, Kyoto 606-8501, Japan
Department of Molecular Biology and Biochemistry, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka, Japan.81-6-6879-341981-6-6879-3410

ytakai{at}molbio.med.osaka-u.ac

We have found a new cell–cell adhesion system at cadherin-based cell–cell adherens junctions (AJs) consisting of at least nectin and l-afadin. Nectin is a Ca2+-independent homophilic immunoglobulin-like adhesion molecule, and l-afadin is an actin filament-binding protein that connects the cytoplasmic region of nectin to the actin cytoskeleton. Both the trans-interaction of nectin and the interaction of nectin with l-afadin are necessary for their colocalization with E-cadherin and catenins at AJs. Here, we examined the mechanism of interaction between these two cell–cell adhesion systems at AJs by the use of {alpha}-catenin–deficient F9 cell lines and cadherin-deficient L cell lines stably expressing their various components. We showed here that nectin and E-cadherin were colocalized through l-afadin and the COOH-terminal half of {alpha}-catenin at AJs. Nectin trans-interacted independently of E-cadherin, and the complex of E-cadherin and {alpha}- and β-catenins was recruited to nectin-based cell–cell adhesion sites through l-afadin without the trans-interaction of E-cadherin. Our results indicate that nectin and cadherin interact through their cytoplasmic domain–associated proteins and suggest that these two cell–cell adhesion systems cooperatively organize cell–cell AJs.

Key Words: immunoglobulin superfamily • afadin • ponsin • catenin • cell–cell adherens junctions

© 2000 The Rockefeller University Press

Abbreviations used in this paper: aa, amino acid(s); Ab, antibody; AJ, adherens junction; F-actin, actin filament; gD, glycoprotein D; GFP, green fluorescent protein; GST, glutathione-S-transferase; HA, hemagglutinin; His6, hexa-histidine; HSV1, herpes simplex virus type 1; Ig, immunoglobulin; TJ, tight junction.


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