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  • Original Paper
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Phosphorylation of the EWS IQ domain regulates transcriptional activity of the EWS/ATF1 and EWS/FLI1 fusion proteins

Oncogene volume 20, pages 1756–1764 (2001)Cite this article

Abstract

Specific chromosomal translocations are commonly present in mesenchymal tumors and frequently involve genes encoding transcription factors. The combination of different domains from unrelated genes results in chimeric proteins believed to play a key role in the neoplastic process. The EWS/ATF1 and EWS/FLI1 fusion proteins associated with Clear Cell Sarcoma and Ewing's Sarcoma, respectively, were utilized to study the comparative effect of the EWS component on two different DNA binding partners. A potential regulatory site within the EWS IQ domain at serine266 was identified, and studies were performed to demonstrate that EWS is phosphorylated in cells and phosphorylation of serine266 regulates transcriptional activity. Mutational analysis showed that elimination of phosphorylation significantly reduced DNA binding activity by EMSA and reporter activation in luciferase assays, whereas phosphorylation mimicry resulted in a partial restoration to wild-type levels. Phosphorylation was also observed to mediate cellular compartmentalization. These studies confirm that IQ domain phosphorylation regulates the transcriptional activity of exogenous EWS/ATF1 and EWS/FLI1 and suggests that post-translational modifications may potentiate the neoplastic behavior of fusion proteins in general. Since the IQ domain is incorporated into only a subset of fusion transcripts, these findings may provide insight into the molecular mechanism underlying clinical heterogeneity observed in Ewing's sarcoma.

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References

  • Bailly RA, Bosselut R, Zucman J, Cormier F, Delattre O, Roussel M, Thomas G, Ghysdael J . 1994 Mol. Cell. Biol. 14: 3230–3241

  • Bassuk AG, Leiden JM . 1997 Adv. Immunol. 64: 65–104

  • Bertolotti A, Melot T, Ackler J, Vigneron M, Delattre O, Tora L . 1998 Mol. Cell. Biol. 18: 1489–1497

  • Bosilevac JM, Gilchrist CA, Jankowski PE, Paul S, Rees AR, Hinrichs SH . 1998 J. Biol. Chem. 273: 16874–16879

  • Bosilevac JM, Olsen RJ, Bridge JA, Hinrichs SH . 1999 J. Biol. Chem. 274: 34811–34818

  • Bridge J, Borek D, Neff J, Huntrakoon M . 1990 Am. J. Clin. Pathol. 93: 26–31

  • Bridge J, Sreekantaiah C, Neff J, Sandberg A . 1991 Cancer Genet. Cytogenet 52: 101–106

  • Bridge JA . 1996 Curr. Opin. Oncol. 8: 284–288

  • Caponigro F, French RC, Kaye SB . 1997 Anticancer Drugs 8: 26–33

  • de Alava E, Kawai A, Healey JH, Fligman I, Meyers PA, Huvos AG, Gerald WL, Jhanwar SC, Argani P, Antonescu CR Pardo-Mindan FJ, Ginsberg J, Womer R, Lawlor ER, Wunder J, Andrulis I, Sorensen PH, Barr FG, Ladanyi M . 1998 J. Clin. Oncol. 16: 1248–1255

  • Deloulme J, Prichard L, Delattre O, Storm D . 1997 J. Biol. Chem. 272: 27369–27377

  • Dohjima T, Ohno T, Banno Y, Nozawa Y, Wen-yi Y, Shimizu K . 2000 Br. J. Cancer 82: 16–19

  • Fontes JD, Strawhecker JM, Bills ND, Lewis RE, Hinrichs SH . 1993 J. Virol. 67: 4436–4441

  • Fujimura Y, Ohno T, Siddique H, Lee L, Rao VN, Reddy ES . 1996 Oncogene 12: 159–167

  • Gilchrist C, Orten D, Bosilevac J, Sanderson S, Hinrichs S . 1995a J. Biochem. Sci. 3: 96–127

  • Gilchrist C, Orten D, Bosilevac J, Sanderson S, Hinrichs S . 1995b Antibody Immunoconj. Radiopharm. 8: 281–298

  • Gonzalez GA, Montminy MR . 1989 Cell 59: 675–680

  • Hunter T, Karin M . 1992 Cell 70: 375–387

  • Jean D, Tellez C, Huang S, Davis DW, Bruns CJ, McConkey DJ, Hinrichs SH, Bar-Eli M . 2000 Oncogene 19: 2721–2730

  • Johnson TK, Schweppe RE, Septer J, Lewis RE . 1999 J. Biol. Chem. 274: 36741–36749

  • Ladnyi M . 1995 Diagn. Molec. Path. 4: 162–173

  • Lam PY, Sublett J, Hollenbach A, Roussel M . 1999 Mol. Cell. Biol. 19: 594–601

  • Li F, Lee K . 1998 Oncogene 16: 1325–1331

  • Lin PP, Brody RI, Hamelin AC, Bradner JE, Healey JH, Ladanyi M . 1999 Cancer Res. 59: 1428–1432

  • May WA, Gishizky ML, Lessnick SL, Lunsford LB, Lewis BC, Delattre O, Zucman J, Thomas G, Denny CT . 1993 Proc. Natl. Acad. Sci. USA 90: 5752–5756

  • Orten DJ, Strawhecker JM, Sanderson SD, Huang D, Prystowsky MB, Hinrichs SH . 1994 J. Biol. Chem. 269: 32254–32263

  • Pan S, Ming K, Dunn T, Li K, Kee K . 1998 Oncogene 16: 1625–1631

  • Peterman R, Mossier B, Aryee DN, Khazak V, Golemis E, Kovar H . 1998 Oncogene 17: 603–610

  • Sarkar G, Sommer SS . 1990 Biotechniques 8: 404–407

  • Zhao L, Giam C . 1992 Proc. Natl. Acad. Sci. USA 89: 7070–7074

  • Zoubek A, Pfleiderer C, Salzer-Kuntschik M, Amann G, Windhager R, Fink FM, Koscielniak E, Delattre O, Strehl S, Ambros PF, Gadner H, Kovar H . 1994 Br. J. Cancer 70: 908–913

  • Zoubek A, Dockhorn-Dworniczak B, Delattre O, Christiansen H, Niggli F, Gatterer-Menz I, Smith TL, Jurgens H, Gadner H, Kovar H . 1996 J. Clin. Oncol. 14: 1245–1251

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Acknowledgements

We thank Robert Lewis for helpful comments on the manuscript, Dee Volle for technical assistance and Marc Ladanyi for providing plasmids pEF7-6 and pS2. This work was supported in part by a grant from the Nebraska Cancer Research Program LB506.

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  1. Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, 69198-6495, Nebraska, NE, USA

    R J Olsen & S H Hinrichs

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  1. R J Olsen
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Olsen, R., Hinrichs, S. Phosphorylation of the EWS IQ domain regulates transcriptional activity of the EWS/ATF1 and EWS/FLI1 fusion proteins. Oncogene 20, 1756–1764 (2001). https://doi.org/10.1038/sj.onc.1204268

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