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J. Biol. Chem., Vol. 279, Issue 29, 30715-30721, July 16, 2004

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Tight Junction Protein Par6 Interacts with an Evolutionarily Conserved Region in the Amino Terminus of PALS1/Stardust^*

Qian Wang, Toby W. Hurd, and Ben Margolis, Investigator of the Howard Hughes Medical Institute¶||

From the Department of Biological Chemistry, the Howard Hughes Medical Institute, and the ^¶Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan 48109-2216

Tight junctions are the structures in mammalian epithelial cells that separate the apical and basolateral membranes and may also be important in the establishment of cell polarity. Two evolutionarily conserved multiprotein complexes, Crumbs-PALS1 (Stardust)-PATJ and Cdc42-Par6-Par3-atypical protein kinase C, have been implicated in the assembly of tight junctions and in polarization of Drosophila melanogaster epithelia. These two complexes have been linked physically and functionally by an interaction between PALS1 and Par6. Here we identify an evolutionarily conserved region in the amino terminus of PALS1 as the Par6 binding site and identify valine and aspartic acid residues in this region as essential for interacting with the PDZ domain of Par6. We have also characterized, in more detail, the amino terminus of Drosophila Stardust and demonstrate that the interaction mechanism between Stardust and Drosophila Par6 is evolutionarily conserved. Par6 interferes with PATJ in binding PALS1, and these two interactions do not appear to function synergistically. Taken together, these results define the molecular mechanisms linking two conserved polarity complexes.

Received for publication, February 22, 2004 , and in revised form, May 3, 2004.

^* This work was supported in part by NIDDK, National Institutes of Health, Grant DK-58208. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed. Tel.: 734-764-3567; Fax: 734-615-4356; E-mail: bmargoli{at}umich.edu.

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