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O-GlcNAcylation is involved in the transcriptional activity of EWS-FLI1 in Ewing's sarcoma

Oncogene volume 28, pages 1280–1284 (2009)Cite this article

Abstract

The oncogene EWS-FLI1 encodes a chimeric transcription factor expressed in Ewing's sarcoma family tumors (ESFTs). EWS-FLI1 target gene expression is thought to drive ESFT pathogenesis and, therefore, inhibition of EWS-FLI1 activity holds high therapeutic promise. As the activity of many transcription factors is regulated by post-translational modifications, we studied the presence of modifications on EWS-FLI1. The immuno-purified fusion-protein was recognized by an antibody specific for O-linked β-N-acetylglucosaminylation, and bound readily to a phosphoprotein-specific dye. Inhibition of Ser/Thr-specific phophatases increased EWS-FLI1 molecular weight and reduced its O-GlcNAc content, suggesting that phosphorylation and O-GlcNAcylation of EWS-FLI1 interact dynamically. By mutation analysis, O-GlcNAcylation was delineated to Ser/Thr residues of the amino-terminal EWS transcriptional-activation domain. Metabolic inhibition of the hexosamine biosynthetic pathway abrogated O-GlcNAcylation of EWS-FLI1 and interfered specifically with transcriptional activation of the EWS-FLI1 target Id2. These results suggest that drugs modulating glycosylation of EWS-FLI1 interfere functionally with its activity and might, therefore, constitute promising additions to the current ESFT chemotherapy.

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Acknowledgements

This study was supported in part by Grant P18046-B12 of the Austrian Science Fund (FWF).

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Authors and Affiliations

  1. Children's Cancer Research Institute, St Anna Kinderkrebsforschung, Vienna, Austria

    R Bachmaier, D N T Aryee, G Jug, M Kauer, M Kreppel & H Kovar

  2. Department of Biology, Hong Kong University of Science and Technology, Kowloon, Hong Kong, SAR, China

    K A Lee

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  1. R Bachmaier
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  2. D N T Aryee
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  3. G Jug
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  4. M Kauer
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  5. M Kreppel
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  6. K A Lee
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  7. H Kovar
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Correspondence to H Kovar.

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Bachmaier, R., Aryee, D., Jug, G. et al. O-GlcNAcylation is involved in the transcriptional activity of EWS-FLI1 in Ewing's sarcoma. Oncogene 28, 1280–1284 (2009). https://doi.org/10.1038/onc.2008.484

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