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Published 19 July 2004. doi:10.1083/jcb.200401078
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 166, Number 2, 237-248
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Endocytosis of E-cadherin regulated by Rac and Cdc42 small G proteins through IQGAP1 and actin filaments

Genkichi Izumi1, Toshiaki Sakisaka1, Takeshi Baba1, Shintaro Tanaka1, Koji Morimoto2, and Yoshimi Takai1

1 Department of Molecular Biology and Biochemistry, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita, Osaka 565-0871, Japan
2 KAN Research Institute Inc., Kyoto 600-8815, Japan

Address correspondence to Yoshimi Takai, Dept. of Molecular Biology and Biochemistry, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita, Osaka 565-0871, Japan. Tel.: 81-6-6879-3410. Fax: 81-6-6879-3419. email: ytakai{at}molbio.med.osaka-u.ac.jp

E-cadherin is a key cell–cell adhesion molecule at adherens junctions (AJs) and undergoes endocytosis when AJs are disrupted by the action of extracellular signals. To elucidate the mechanism of this endocytosis, we developed here a new cell-free assay system for this reaction using the AJ-enriched fraction from rat liver. We found here that non-trans-interacting, but not trans-interacting, E-cadherin underwent endocytosis in a clathrin-dependent manner. The endocytosis of trans-interacting E-cadherin was inhibited by Rac and Cdc42 small G proteins, which were activated by trans-interacting E-cadherin or trans-interacting nectins, which are known to induce the formation of AJs in cooperation with E-cadherin. This inhibition was mediated by reorganization of the actin cytoskeleton by Rac and Cdc42 through IQGAP1, an actin filament-binding protein and a downstream target of Rac and Cdc42. These results indicate the important role of the Rac/Cdc42-IQGAP1 system in the dynamic organization and maintenance of the E-cadherin–based AJs.

Key Words: cell-free assay; E-cadherin; endocytosis; Rho family small G proteins; IQGAP1


Abbreviations used in this paper: AJ, adherens junction; CAMs, cell–cell adhesion molecules; CCV, clathrin-coated vesicle; HSP, high-speed pellet; Lat-A, latrunculin A; MSP, medium-speed pellet; MSS, medium-speed supernatant.


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