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Vol. 15, Issue 4, 1981-1990, April 2004
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University of Michigan, Department of Biological Chemistry, Ann Arbor, Michigan 48109;
University of Michigan, Department of Internal Medicine, Ann Arbor, Michigan 48109; and
* University of Michigan, Howard Hughes Medical Institute, Ann Arbor, Michigan 48109
Submitted August 22, 2003;
Revised November 30, 2003;
Accepted December 8, 2003
Monitoring Editor: Mark Ginsberg
Prior work in our laboratory established a connection between the PALS1/PATJ/CRB3 and Par6/Par3/aPKC protein complexes at the tight junction of mammalian epithelial cells. Utilizing a stable small interfering RNA expression system, we have markedly reduced expression of the tight junction-associated protein PALS1 in MDCKII cells. The loss of PALS1 resulted in a corresponding loss of expression of PATJ, a known binding partner of PALS1, but had no effect on the expression of CRB3. However, the absence of PALS1 and PATJ expression did result in the decreased association of CRB3 with members of the Par6/Par3/aPKC protein complex. The consequences of the loss of PALS1 and PATJ were exhibited by a delay in the polarization of MDCKII monolayers after calcium switch, a decrease in the transepithelial electrical resistance, and by the inability of these cells to form lumenal cysts when grown in a collagen gel matrix. These defects in polarity determination may be the result of the lack of recruitment of aPKC to the tight junction in PALS1-deficient cells, as observed by confocal microscopy, and subsequent alterations in downstream signaling events.
Corresponding author. E-mail address: bmargoli{at}umich.edu.
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