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Originally published In Press as doi:10.1074/jbc.M201177200 on May 20, 2002

J. Biol. Chem., Vol. 277, Issue 30, 27501-27509, July 26, 2002
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The Carboxyl Terminus of Zona Occludens-3 Binds and Recruits a Mammalian Homologue of Discs Lost to Tight Junctions*

Michael H. RohDagger §, Chia-Jen Liu, Stephanie Laurinec||, and Ben MargolisDagger ||**

From the || Howard Hughes Medical Institute, Departments of Dagger  Biological Chemistry and  Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan 48109

Mammalian homologues of the Drosophila polarity proteins Stardust, Discs Lost, and Crumbs have been identified as Pals1, Pals1-associated tight junction protein (PATJ), and human Crumbs homologue 1 (CRB1), respectively. We have previously demonstrated that PATJ, Pals1, and CRB1 can form a tripartite tight junction complex in epithelial cells and that PATJ recruits Pals1 to tight junctions. Here, we observed that the Pals1/PATJ interaction was not crucial for the ultimate targeting of PATJ itself to tight junctions. This prompted us to examine if any of the 10 post-synaptic density-95/Discs Large/zona occludens-1 (PDZ) domains of PATJ could bind to the carboxyl termini of known tight junction constituents. We found that the 6th and 8th PDZ domains of PATJ can interact with the carboxyl termini of zona occludens-3 (ZO-3) and claudin 1, respectively. PATJ missing the 6th PDZ domain was found to mislocalize away from cell contacts. Surprisingly, deleting the 8th PDZ domain had little effect on PATJ localization. Finally, reciprocal co-immunoprecipitation experiments revealed that full-length ZO-3 can associate with PATJ. Hence, the PATJ/ZO-3 interaction is likely important for recruiting PATJ and its associated proteins to tight junctions.

* This work was supported in part by NIDDK, National Institutes of Health Grant DK58208.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Supported by the Medical Scientist Training Program Grant T32 GM07863 and Predoctoral Genetics Training Program Grant T32 GM07544-24 to the University of Michigan during the course of these studies.

** Investigator of the Howard Hughes Medical Institute. To whom corresponding should be addressed: Howard Hughes Medical Inst., University of Michigan Medical Center, 4570 MSRB II, Box 0650, 1150 W. Medical Center Dr., Ann Arbor, MI 48109-0650. Tel.: 734-764-3567; Fax: 734-763-9323; E-mail: bmargoli@umich.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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