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Published online March 12, 2007
doi:10.1083/jcb.200612080
The Journal of Cell Biology, Vol. 176, No. 6, 779-786
The Rockefeller University Press, 0021-9525 $30.00
© 2007 Ikenouchi et al.
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 Requirement of ZO-1 for the formation of belt-like adherens junctions during epithelial cell polarization

Junichi Ikenouchi1, Kazuaki Umeda3, Sachiko Tsukita1,2,5, Mikio Furuse4, and Shoichiro Tsukita1,5

1 Department of Cell Biology and 2 Division of Health Sciences, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan
3 Department of Molecular Pharmacology, Graduate School of Medical Sciences, Kumamoto University, Honjo, Kumamoto 860-8556, Japan
4 Division of Cellular and Molecular Medicine, Kobe University Graduate School of Medicine, Chuo-ku, Kobe 650-0017, Japan
5 Solution Oriented Research for Science and Technology, Japan Science and Technology Corporation, Sakyo-ku, Kyoto 606-8501, Japan

Correspondence to Junichi Ikenouchi: ikenouti{at}mfour.med.kyoto-u.ac.jp; or Sachiko Tsukita: atsukita{at}mfour.med.kyoto-u.ac.jp

The molecular mechanisms of how primordial adherens junctions (AJs) evolve into spatially separated belt-like AJs and tight junctions (TJs) during epithelial polarization are not well understood. Previously, we reported the establishment of ZO-1/ZO-2–deficient cultured epithelial cells (1[ko]/2[kd] cells), which lacked TJs completely. In the present study, we found that the formation of belt-like AJs was significantly delayed in 1(ko)/2(kd) cells during epithelial polarization. The activation of Rac1 upon primordial AJ formation is severely impaired in 1(ko)/2(kd) cells. Our data indicate that ZO-1 plays crucial roles not only in TJ formation, but also in the conversion from "fibroblastic" AJs to belt-like "polarized epithelial" AJs through Rac1 activation. Furthermore, to examine whether ZO-1 itself mediate belt-like AJ and TJ formation, respectively, we performed a mutational analysis of ZO-1. The requirement for ZO-1 differs between belt-like AJ and TJ formation. We propose that ZO-1 is directly involved in the establishment of two distinct junctional domains, belt-like AJs and TJs, during epithelial polarization.

Dr. Shoichiro Tsukita died on 11 December 2005.

Abbreviations used in this paper: AJ, adherens junction; DA, dominant-active; MAGUK, membrane-associated guanlyate kinase; TJ, tight junction.


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