title: Phosphorylation state regulates the localization of Scribble at adherens junctions and its association with E-cadherin-catenin complexes.
abstract: Mammalian ortholog of Scribble tumor suppressor has been reported to regulate cadherin-mediated epithelial cell adhesion by stabilizing the coupling of E-cadherin with catenins, but the molecular mechanism involved remains unknown. In this study, we investigated the relationship between the localization of mouse Scribble at cadherin-based adherens junctions (AJs) and its phosphorylation state. Immunofluorescence staining confirmed that Scribble was localized at AJs as well as at the basolateral plasma membrane in epithelial cells. We found that Scribble was detected as two bands by Western blotting analysis and that the band shift to the higher molecular weight was dependent on its phosphorylation at Ser 1601. Triton X-100 treatment extracted Scribble localized on the basolateral membrane but not Scribble localized at AJs in cultured epithelial cells, and the Triton X-100-resistant Scribble was the Ser 1601-unphosphorylated form. Conversely, an in-house-generated antibody that predominantly recognized Ser 1601-phosphorylated Scribble only detected Scribble protein on the lateral plasma membrane. Furthermore, Ser 1601-unphosphorylated Scribble was selectively coprecipitated with E-cadherin-catenin complexes in E-cadherin-expressing mouse L fibroblasts. Taken together, these results suggest that the phosphorylation state of Scribble regulates its complex formation with the E-cadherin-catenin system and may control cadherin-mediated cell-cell adhesion.
authors.affiliation: Division of Cell Biology, Department of Physiology and Cell Biology, Kobe University Graduate School of Medicine, Kobe 650-0017, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Yoshihara
authors.forename: Junichi
authors.initials: J
authors.lastname: Ikenouchi
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Izumi
authors.forename: Masaya
authors.initials: M
authors.lastname: Akashi
authors.forename: Shoichiro
authors.initials: S
authors.lastname: Tsukita
authors.forename: Mikio
authors.initials: M
authors.lastname: Furuse
mesh_terms: D022005:Adherens Junctions / Q000737:chemistry*; D000818:Animals; D015820:Cadherins / Q000378:metabolism*; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D004847:Epithelial Cells; D005347:Fibroblasts; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis / Q000378:metabolism*; D051379:Mice; D010766:Phosphorylation; D011485:Protein Binding; D051178:alpha Catenin / Q000378:metabolism*
chemical_list: D015820:Cadherins; D047908:Intracellular Signaling Peptides and Proteins; D051178:alpha Catenin; C508433:scrib protein, mouse