Phosphorylation state regulates the localization of Scribble at adherens junctions and its association with E-cadherin-catenin complexes

Ken Yoshihara, Junichi Ikenouchi, Yasushi Izumi, Masaya Akashi, Shoichiro Tsukita, Mikio Furuse

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Pages (from-to)413-422
Number of pages10
JournalExperimental Cell Research
Volume317
Issue number4
DOIs
Publication statusPublished - Feb 15 2011

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Adherens Junctions
Catenins
Cadherins
Phosphorylation
Epithelial Cells
Octoxynol
Cell Adhesion
Cell Membrane
Fluorescent Antibody Technique
Cultured Cells
Fibroblasts
Molecular Weight
Western Blotting
Staining and Labeling
Membranes
Antibodies

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

Phosphorylation state regulates the localization of Scribble at adherens junctions and its association with E-cadherin-catenin complexes. / Yoshihara, Ken; Ikenouchi, Junichi; Izumi, Yasushi; Akashi, Masaya; Tsukita, Shoichiro; Furuse, Mikio.

In: Experimental Cell Research, Vol. 317, No. 4, 15.02.2011, p. 413-422.

Research output: Contribution to journalArticle

Yoshihara, Ken ; Ikenouchi, Junichi ; Izumi, Yasushi ; Akashi, Masaya ; Tsukita, Shoichiro ; Furuse, Mikio. / Phosphorylation state regulates the localization of Scribble at adherens junctions and its association with E-cadherin-catenin complexes. In: Experimental Cell Research. 2011 ; Vol. 317, No. 4. pp. 413-422.
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