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First published online 27 May 2003
doi: 10.1242/jcs.00500
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Research Article |
1 Department of Biological Chemistry
2 Howard Hughes Medical Institute
3 Department of Internal Medicine, University of Michigan Medical School, Ann
Arbor, MI 48109-0650, USA
Author for correspondence (e-mail:
bmargoli{at}umich.edu)
Accepted 24 March 2003
In Drosophila, the Crumbs–Stardust–Discs-lost complex is required during the establishment of polarized epithelia. Embryos that lack a component of this complex or overexpress Crumbs exhibit defects in epithelial morphogenesis. We recently cloned a novel mammalian epithelial Crumbs isoform, Crumbs3 (CRB3). CRB3 exists in a complex at tight junctions (TJs) with Pals1 and PATJ, the mammalian homologues of Stardust and Discs lost, respectively. Here, we observe that overexpression of CRB3 leads to delayed TJ formation in MDCK epithelial cell monolayers and disruption of polarity in MDCK cysts cultured in collagen. Both phenomena require the last four residues of CRB3. Next, we expressed, in MDCK cells, a dominant-negative Myc-Lin-2–Pals1 chimeric protein, where the PDZ domain of Lin-2 was replaced with that of Pals1. TJ and apical polarity defects were also observed in these cells. Collectively, this suggests that the CRB-Pals1 interaction is important for formation of TJs and polarized epithelia. These results provide insight into the function of the mammalian Crumbs complex during TJ formation and epithelial polarization.
Key words: Crumbs3, Pals1, PDZ domain, Tight junction, Polarity
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