Direct association of occludin with ZO-1 and its possible involvement in the localization of occludin at tight junctions

doi:10.1083/jcb.127.6.1617

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Direct association of occludin with ZO-1 and its possible involvement in the localization of occludin at tight junctions

M Furuse, M Itoh, T Hirase, A Nagafuchi, S Yonemura, S Tsukita and S Tsukita
Department of Information Physiology, National Institute for Physiological Sciences, Okazaki, Aichi, Japan.

Occludin is an integral membrane protein localizing at tight junctions (TJ) with four transmembrane domains and a long COOH-terminal cytoplasmic domain (domain E) consisting of 255 amino acids. Immunofluorescence and laser scan microscopy revealed that chick full- length occludin introduced into human and bovine epithelial cells was correctly delivered to and incorporated into preexisting TJ. Further transfection studies with various deletion mutants showed that the domain E, especially its COOH-terminal approximately 150 amino acids (domain E358/504), was necessary for the localization of occludin at TJ. Secondly, domain E was expressed in Escherichia coli as a fusion protein with glutathione-S-transferase, and this fusion protein was shown to be specifically bound to a complex of ZO-1 (220 kD) and ZO-2 (160 kD) among various membrane peripheral proteins. In vitro binding analyses using glutathione-S-transferase fusion proteins of various deletion mutants of domain E narrowed down the sequence necessary for the ZO-1/ZO-2 association into the domain E358/504. Furthermore, this region directly associated with the recombinant ZO-1 produced in E. coli. We concluded that occludin itself can localize at TJ and directly associate with ZO-1. The coincidence of the sequence necessary for the ZO-1 association with that for the TJ localization suggests that the association with underlying cytoskeletons through ZO-1 is required for occludin to be localized at TJ.
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Andreeva, A. Y., Krause, E., Muller, E.-C., Blasig, I. E., Utepbergenov, D. I. (2001). Protein Kinase C Regulates the Phosphorylation and Cellular Localization of Occludin. J. Biol. Chem. 276: 38480-38486 [Abstract] [Full Text]
Defamie, N., Mograbi, B., Roger, C., Cronier, L., Malassine, A., Brucker-Davis, F., Fenichel, P., Segretain, D., Pointis, G. (2001). Disruption of gap junctional intercellular communication by lindane is associated with aberrant localization of connexin43 and zonula occludens-1 in 42GPA9 Sertoli cells. Carcinogenesis 22: 1537-1542 [Abstract] [Full Text]
Roche, H M, Terres, A M, Black, I B, Gibney, M J, Kelleher, D (2001). Fatty acids and epithelial permeability: effect of conjugated linoleic acid in Caco-2 cells. Gut 48: 797-802 [Abstract] [Full Text]
Atkinson, K. J., Rao, R. K. (2001). Role of protein tyrosine phosphorylation in acetaldehyde-induced disruption of epithelial tight junctions. Am. J. Physiol. Gastrointest. Liver Physiol. 280: G1280-G1288 [Abstract] [Full Text]
Tiwari-Woodruff, S. K., Buznikov, A. G., Vu, T. Q., Micevych, P. E., Chen, K., Kornblum, H. I., Bronstein, J. M. (2001). Osp/Claudin-11 Forms a Complex with a Novel Member of the Tetraspanin Super Family and {beta}1 Integrin and Regulates Proliferation and Migration of Oligodendrocytes. JCB 153: 295-306 [Abstract] [Full Text]
Kojima, T., Kokai, Y., Chiba, H., Osanai, M., Kuwahara, K., Mori, M., Mochizuki, Y., Sawada, N. (2001). Occludin and Claudin-1 Concentrate in the Midbody of Immortalized Mouse Hepatocytes During Cell Division. J. Histochem. Cytochem. 49: 333-340 [Abstract] [Full Text]
Nusrat, A., von Eichel-Streiber, C., Turner, J. R., Verkade, P., Madara, J. L., Parkos, C. A. (2001). Clostridium difficile Toxins Disrupt Epithelial Barrier Function by Altering Membrane Microdomain Localization of Tight Junction Proteins. Infect. Immun. 69: 1329-1336 [Abstract] [Full Text]
Marzioni, D., Banita, M., Felici, A., Paradinas, F.J., Newlands, E., De Nictolis, M., Muhlhauser, J., Castellucci, M. (2001). Expression of ZO-1 and occludin in normal human placenta and in hydatidiform moles. Mol Hum Reprod 7: 279-285 [Abstract] [Full Text]
Decaens, C., Cassio, D. (2001). Spatiotemporal expression of catenins, ZO-1, and occludin during early polarization of hepatic WIF-B9 cells. Am. J. Physiol. Cell Physiol. 280: C527-C539 [Abstract] [Full Text]