title: Recruitment of E-cadherin associated with alpha- and beta-catenins and p120ctn to the nectin-based cell-cell adhesion sites by the action of 12-O-tetradecanoylphorbol-13-acetate in MDCK cells.
abstract: The formation of tight junctions (TJs) is dependent on the formation of adherens junctions (AJs) in MDCK cells. E-Cadherin and nectin are major cell-cell adhesion molecules (CAMs) at AJs, whereas claudin, occludin and junctional adhesion molecule (JAM) are major CAMs at TJs. When MDCK cells precultured at 2 microm Ca(2+) are cultured at 2 mm Ca(2+), nectin first forms cell-cell adhesion and recruits E-cadherin to the nectin-based cell-cell adhesion sites to form AJs. Thereafter, nectin recruits first JAM-A and then claudin-1 and occludin to the apical side of AJs to form TJs. In contrast, when MDCK cells precultured at 2 microm Ca(2+) are cultured at 2 microm Ca(2+) in the presence of a phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), a TJ-like structure is formed without the formation of the E-cadherin-based AJs. We showed here that GFP-E-cadherin, which did not trans-interact due to 2 microm Ca(2+) but associated with alpha- and beta-catenins and p120(ctn), was recruited to the nectin-based cell-cell adhesion sites by the action of TPA. The nectin inhibitors, which inhibited the trans-interaction of nectin, inhibited the recruitment of GFP-E-cadherin and their associating catenins by the action of TPA. Microbeads coated with the extracellular fragment of nectin recruited not only cellular nectin but also GFP-E-cadherin and their associating catenins by the action of TPA. These results indicate that when the TJ-like structure is formed by the action of TPA, non-trans-interacting E-cadherin and its associating catenins are recruited to the nectin-based cell-cell adhesion sites and that the trans-interaction of E-cadherin is not essential for the formation of TJs.
authors.affiliation: Department of Molecular Biology and Biochemistry, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Japan.
authors.forename: Ryoko
authors.initials: R
authors.lastname: Okamoto
authors.forename: Kenji
authors.initials: K
authors.lastname: Irie
authors.forename: Akio
authors.initials: A
authors.lastname: Yamada
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Katata
authors.forename: Atsunori
authors.initials: A
authors.lastname: Fukuhara
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Takai
mesh_terms: D000818:Animals; D015820:Cadherins / Q000378:metabolism*; D051177:Catenins; D002448:Cell Adhesion / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000276:immunology / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D004285:Dogs; D007074:Immunoglobulin G; D008863:Microspheres; D000075983:Nectins; D010750:Phosphoproteins / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000378:metabolism*; D019108:Tight Junctions / Q000502:physiology; D015534:Trans-Activators / Q000378:metabolism*; D051178:alpha Catenin; D051176:beta Catenin; D000095002:Delta Catenin
chemical_list: D015820:Cadherins; D051177:Catenins; D015815:Cell Adhesion Molecules; D003598:Cytoskeletal Proteins; D007074:Immunoglobulin G; D000075983:Nectins; D010750:Phosphoproteins; D015534:Trans-Activators; D051178:alpha Catenin; D051176:beta Catenin; D013755:Tetradecanoylphorbol Acetate; D000095002:Delta Catenin