Snail Destabilizes Cell Surface Crumbs3a

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Abstract
During epithelial to mesenchymal transition (EMT), cells modulate expression of proteins resulting in loss of apical‐basal polarity. Effectors of this EMT switch target the polarity protein Crumbs3a, a small transmembrane protein that is essential for generation of the apical membrane and tight junctions of mammalian epithelial cells. We previously showed that the Crumbs3 gene is a direct target of transcriptional regulation by Snail, a potent inducer of EMT. However, Snail has also been shown to have multiple non‐transcriptional roles, including regulation of cell adhesion, proliferation and survival. Using SNAP‐tag labeling, we determined that cell surface Crumbs3a has a half‐life of approximately 3 h and that this cell surface half‐life is significantly reduced when EMT is induced by Snail. We further observe that Snail induces differential glycosylation of Crumbs3a, including sialylation, suggesting a mechanism by which Crumbs3a may be destabilized. These results indicate that Crumbs3a is a post‐translational target of Snail, in addition to being a transcriptional target. We conclude that Snail's ability to post‐translationally modify and destabilize Crumbs3a augments the depolarizing process of EMT.

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