Adipogenesis stimulates the nuclear localization of EWS with an increase in its O-GlcNAc glycosylation in 3T3-L1 cells |
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Authors: | Qiang Li Kazuo Kamemura |
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Affiliation: | Department of Bioscience, Nagahama Institute of Bio-Science and Technology, 1266 Tamura, Nagahama, Shiga 526-0829, Japan |
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Abstract: | Although the Ewing sarcoma (EWS) proto-oncoprotein is found in the nucleus and cytosol and is associated with the cell membrane, the regulatory mechanisms of its subcellular localization are still unclear. Here we found that adipogenic stimuli induce the nuclear localization of EWS in 3T3-L1 cells. Tyrosine phosphorylation in the C-terminal PY-nuclear localization signal of EWS was negative throughout adipogenesis. Instead, an adipogenesis-dependent increase in O-linked β-N-acetylglucosamine (O-GlcNAc) glycosylation of EWS was observed. Pharmacological inactivation of O-GlcNAcase in preadipocytes promoted perinuclear localization of EWS. Our findings suggest that the nuclear localization of EWS is partly regulated by the glycosylation. |
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Keywords: | DMEM, Dulbecco&rsquo s modified Eagle&rsquo s medium EWS, Ewing sarcoma proto-oncoprotein HRP, horseradish peroxidase NLS, nuclear localization signal O-GlcNAc, O-linked β-N-acetylglucosamine PBS, phosphate-buffered saline |
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