PAR1b Promotes Cell-Cell Adhesion and Inhibits Dishevelled-mediated Transformation of MDCK Cells

Authors:
Elbert M, Cohen D, Musch A
In:
Source: Mol Biol Cell
Publication Date: (2006)
Issue: 17(8): 3345-55
Research Area:
Cancer Research/Cell Biology
Cells used in publication:
MDCK
Species: canine
Tissue Origin: kidney
Platform:
Nucleofector® I/II/2b
Experiment
Nucleofection of 10-15µg pSuperior vector (inducable shRNA system) into 4 million cells per sample according to the amaxa optimized protocol for MDCK cells.
Abstract
Mammalian Par1 is a family of serine/threonine kinases comprised of four homologous isoforms that have been associated with tumor suppression and differentiation of epithelial and neuronal cells, yet little is known about their cellular functions. In polarizing kidney epithelial (MDCK) cells, the Par1 isoform Par1b/MARK2/EMK1 promotes the E-Cadherin-dependent compaction, columnarization, and cytoskeletal organization characteristic of differentiated columnar epithelia. Here we identify two functions of Par1b that likely contribute to its role as a tumor suppressor in epithelial cells: (i) The kinase promotes cell-cell adhesion and resistance of E-cadherin to extraction by nonionic detergents, a measure for the association of the E-Cadherin cytoplasmic domain with the actin cytoskeleton, which is critical for E-Cadherin function. (ii) Par1b attenuates the effect of Dishevelled (Dvl)-expression, an inducer of wnt-signaling that causes transformation of epithelial cells. Although Dvl is a known Par1 substrate in vitro, we determined, after mapping the PAR1b-phosphorylation sites in Dvl, that PAR1b did not antagonize Dvl-signaling by phosphorylating the wnt-signaling molecule. Instead, our data suggest that both proteins function antagonistically to regulate the assembly of functional E-cadherin-dependent adhesion complexes.