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Published 13 October 2003. doi:10.1083/jcb.200301075
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© The Rockefeller University Press, 0021-9525/2003/10/165 $8.00
The Journal of Cell Biology, Volume 163, Number 1, 165-176

Article

 CD151 regulates epithelial cell–cell adhesion through PKC- and Cdc42-dependent actin cytoskeletal reorganization

Masaki Shigeta1, Noriko Sanzen1,2, Masayuki Ozawa3, Jianguo Gu1, Hitoshi Hasegawa4 and Kiyotoshi Sekiguchi1,2

1 Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan
2 Sekiguchi Biomatrix Signaling Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Aichi Medical University, Nagakute, Aichi 480-1195, Japan
3 Department of Biochemistry and Molecular Biology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8520, Japan
4 First Department of Internal Medicine, Ehime University School of Medicine, Shigenobu, Ehime 791-0295, Japan

Address correspondence to Kiyotoshi Sekiguchi, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan. Tel.: 81-6-6879-8617. Fax.: 81-6-6879-8619. email: sekiguch{at}protein.osaka-u.ac.jp

CD151, a member of the tetraspanin family proteins, tightly associates with integrin {alpha}3ß1 and localizes at basolateral surfaces of epithelial cells. We found that overexpression of CD151 in A431 cells accelerated intercellular adhesion, whereas treatment of cells with anti-CD151 mAb perturbed the integrity of cortical actin filaments and cell polarity. E-Cadherin puncta formation, indicative of filopodia-based adhesion zipper formation, as well as E-cadherin anchorage to detergent-insoluble cytoskeletal matrix, was enhanced in CD151-overexpressing cells. Levels of GTP-bound Cdc42 and Rac were also elevated in CD151-overexpressing cells, suggesting the role of CD151 in E-cadherin–mediated cell–cell adhesion as a modulator of actin cytoskeletal reorganization. Consistent with this possibility, engagement of CD151 by the substrate-adsorbed anti-CD151 mAb induced prominent Cdc42-dependent filopodial extension, which along with E-cadherin puncta formation, was strongly inhibited by calphostin C, a protein kinase C (PKC) inhibitor. Together, these results indicate that CD151 is involved in epithelial cell–cell adhesion as a modulator of PKC- and Cdc42-dependent actin cytoskeletal reorganization.

Key Words: cell adhesion; tetraspanin; integrin; actin cytoskeleton; Rho family GTPase

Abbreviations used in this paper: cPKC, conventional PKC; dn, dominant-negative; GST-CRIB, a fusion protein of GST to the Cdc42/Rac-interactive–binding domain of PAK1; GST-RB, a fusion protein of GST to the Rho-binding domain of Rho kinase.


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