Loss of PALS1 Expression Leads to Tight Junction and Polarity Defects

Samuel W. Straight, Kunyoo Shin, Vanessa C. Fogg, Shuling Fan, Chia Jen Liu, Michael Roh, Ben Margolis

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)1981-1990
Number of pages10
JournalMolecular Biology of the Cell
Volume15
Issue number4
DOIs
StatePublished - Apr 2004
Externally publishedYes

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Tight Junctions
Tight Junction Proteins
Electric Impedance
Confocal Microscopy
Small Interfering RNA
Cysts
Collagen
Gels
Epithelial Cells
Calcium
PKC-3 protein

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Straight, S. W., Shin, K., Fogg, V. C., Fan, S., Liu, C. J., Roh, M., & Margolis, B. (2004). Loss of PALS1 Expression Leads to Tight Junction and Polarity Defects. Molecular Biology of the Cell, 15(4), 1981-1990. https://doi.org/10.1091/mbc.E03-08-0620

Loss of PALS1 Expression Leads to Tight Junction and Polarity Defects. / Straight, Samuel W.; Shin, Kunyoo; Fogg, Vanessa C.; Fan, Shuling; Liu, Chia Jen; Roh, Michael; Margolis, Ben.

In: Molecular Biology of the Cell, Vol. 15, No. 4, 04.2004, p. 1981-1990.

Research output: Contribution to journalArticle

Straight, SW, Shin, K, Fogg, VC, Fan, S, Liu, CJ, Roh, M & Margolis, B 2004, 'Loss of PALS1 Expression Leads to Tight Junction and Polarity Defects', Molecular Biology of the Cell, vol. 15, no. 4, pp. 1981-1990. https://doi.org/10.1091/mbc.E03-08-0620
Straight, Samuel W. ; Shin, Kunyoo ; Fogg, Vanessa C. ; Fan, Shuling ; Liu, Chia Jen ; Roh, Michael ; Margolis, Ben. / Loss of PALS1 Expression Leads to Tight Junction and Polarity Defects. In: Molecular Biology of the Cell. 2004 ; Vol. 15, No. 4. pp. 1981-1990.
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