Beta-catenin cleavage in non-apoptotic cells with reduced cell adhesion activity.
Beta-catenin functions both as a regulator of cadherin-mediated cell-cell adhesion and a mediator of Wnt signaling. Recently, caspase-3-dependent cleavage of beta-catenin was demonstrated to occur during apoptosis. Here, we show that beta-catenin is proteolytically cleaved in G401 Wilms' tumor cells that were detached from the culture dish. Beta-catenin cleavage products of the same electrophoretic mobility were detected in G401 cells after induction of apoptosis with staurosporine and cell cycle arrest by aphidicolin. The detached cells show no sign of anoikis and approximately 90% of the floating cells were able to reattach to new dishes. Furthermore, beta-catenin was not cleaved in cells cultured on dishes coated with poly(2-hydroxyethylmethacrylate), which inhibits cellular attachment on the dishes, with approximately 90% of cells viable under these conditions. All beta-catenin cleavage products lost N-terminal and C-terminal regions and were unable to associate with alpha-catenin, which is responsible for actin filament binding and organization. However, they were still able to associate with E-cadherin. Aggregation assays revealed that the floating cells had weak aggregation compared with the attached cells. These results suggest that the cleavage of beta-catenin during cell detachment functions at least in part to remove the alpha-catenin-binding domain, thereby reducing cell adhesion activity.
Nakamoto
Kazutaka
K
Department of Biochemistry and Molecular Biology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8544, Japan.
Kuratsu
Jun-Ichi
J
Ozawa
Masayuki
M
D000595:Amino Acid Sequence; D016590:Aphidicolin / Q000494:pharmacology; D017209:Apoptosis; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism*; D009396:Wilms Tumor / Q000473:pathology; D051176:beta Catenin
C495270:CTNNB1 protein, human; D003598:Cytoskeletal Proteins; D004791:Enzyme Inhibitors; D015534:Trans-Activators; D051176:beta Catenin; D016590:Aphidicolin