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- Title
O -GlcNAcylation and Regulation of Galectin-3 in Extraembryonic Endoderm Differentiation.
- Authors
Gatie, Mohamed I.; Spice, Danielle M.; Garha, Amritpal; McTague, Adam; Ahmer, Mariam; Timoshenko, Alexander V.; Kelly, Gregory M.
- Abstract
The regulation of proteins through the addition and removal of O-linked β-N-acetylglucosamine (O-GlcNAc) plays a role in many signaling events, specifically in stem cell pluripotency and the regulation of differentiation. However, these post-translational modifications have not been explored in extraembryonic endoderm (XEN) differentiation. Of the plethora of proteins regulated through O-GlcNAc, we explored galectin-3 as a candidate protein known to have various intracellular and extracellular functions. Based on other studies, we predicted a reduction in global O-GlcNAcylation levels and a distinct galectin expression profile in XEN cells relative to embryonic stem (ES) cells. By conducting dot blot analysis, XEN cells had decreased levels of global O-GlcNAc than ES cells, which reflected a disbalance in the expression of genes encoding O-GlcNAc cycle enzymes. Immunoassays (Western blot and ELISA) revealed that although XEN cells (low O-GlcNAc) had lower concentrations of both intracellular and extracellular galectin-3 than ES cells (high O-GlcNAc), the relative secretion of galectin-3 was significantly increased by XEN cells. Inducing ES cells toward XEN in the presence of an O-GlcNAcase inhibitor was not sufficient to inhibit XEN differentiation. However, global O-GlcNAcylation was found to decrease in differentiated cells and the extracellular localization of galectin-3 accompanies these changes. Inhibiting global O-GlcNAcylation status does not, however, impact pluripotency and the ability of ES cells to differentiate to the XEN lineage.
- Subjects
GALECTINS; ENDODERM; EMBRYONIC stem cells; CELLULAR control mechanisms; IMMUNOASSAY; POST-translational modification
- Publication
Biomolecules (2218-273X), 2022, Vol 12, Issue 5, p623
- ISSN
2218-273X
- Publication type
Article
- DOI
10.3390/biom12050623