title: The extracellular domains of E- and N-cadherin determine the scattered punctate localization in epithelial cells and the cytoplasmic domains modulate the localization.
abstract: The accumulation of classical cadherins is essential for their function, but the mechanism is poorly understood. Hence, we investigated the accumulation of E- and N-cadherin and the formation of cell junctions in epithelial cells. Immunostaining revealed a scattered dot-like accumulation of E- and N-cadherin throughout the lateral membrane in MDCK II and other epithelial cells. Mutant E-cadherin lacking the beta-catenin binding site accumulated granularly at cell-cell contact sites and showed weak cell aggregation activity in cadherin-deficient epithelial cells, MIA PaCa2 cells. Mutant E-cadherin lacking the p120-catenin binding site exhibited scattered punctate accumulation and strong cell adhesion activity in MIA PaCa2 cells. Electron microscopy demonstrated that MIA PaCa2 transfectants of E-cadherin containing beta-catenin binding site formed adherens junction, whereas E-cadherin lacking the binding site did not. Mutant N-cadherins showed accumulation properties similar to those of corresponding mutant E-cadherins. Moreover, wild type and mutant N-cadherin lacking the p120-catenin binding site showed subapical accumulation in polarized DLD-1 cells, whereas mutant N-cadherin lacking beta-catenin binding site did not. These results indicate that the extracellular domains of E- and N-cadherin determines the basic localization pattern, whereas the cytoplasmic domains modulate it thereby affects the cell adhesion activity, subapical accumulation, and the formation of adherens junction.
authors.affiliation: Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, Sanda-shi, Hyogo-ken, Japan.
authors.forename: Chisa
authors.initials: C
authors.lastname: Ozaki
authors.forename: Shuichi
authors.initials: S
authors.lastname: Obata
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Yamanaka
authors.forename: Sachiko
authors.initials: S
authors.lastname: Tominaga
authors.forename: Shintaro T
authors.initials: ST
authors.lastname: Suzuki
mesh_terms: D022005:Adherens Junctions / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D015820:Cadherins / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D002448:Cell Adhesion; D002449:Cell Aggregation; D002460:Cell Line; D003593:Cytoplasm / Q000737:chemistry; D004285:Dogs; D004847:Epithelial Cells / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D051379:Mice; D046529:Microscopy, Electron, Transmission; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D017384:Sequence Deletion; D051176:beta Catenin / Q000172:deficiency
chemical_list: D015820:Cadherins; D051176:beta Catenin