Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Repression of the gene encoding the TGF-β type II receptor is a major target of the EWS-FLI1 oncoprotein

Nature Genetics volume 23, pages 222–227 (1999)Cite this article

A Correction to this article was published on 01 December 1999

Abstract

Chromosomal translocations resulting in the expression of chimaeric transcription factors are frequently observed in tumour cells1, and have been suggested to be a common mechanism in human carcinogenesis. Ewing sarcoma and related peripheral primitive neuroectodermal tumours share recurrent translocations that fuse the gene EWSR1 (formerly EWS) from 22q–12 to FLI1 and genes encoding other ETS transcription factors2,3,4 (which bind DNA through the conserved ETS domain5,6). It has been shown that transduction of the gene EWSR1-FLI1 (encoding EWS-FLI1 protein) can transform NIH3T3 cells, and that mutants containing a deletion in either the EWS domain or the DNA-binding domain in FLI1 lose this ability5,6,7,8. This indicates that the EWS-FLI1 fusion protein may act as an aberrant transcription factor, but the exact mechanism of oncogenesis remains unknown. Because ETS transcription factors regulate expression of TGFBR2 (encoding the TGF-β type II receptor, TGF-β RII; Refs 9,14), a putative tumour suppressor gene, we hypothesized that TGFBR2 may be a target of the EWS-FLI1 fusion protein. We show here that embryonic stem (ES) cell lines with the EWSR1-FLI1 fusion have reduced TGF-β sensitivity, and that fusion-positive ES cells and primary tumours both express low or undetectable levels of TGFBR2 mRNA and protein product. Co-transfection of FLI1 and the TGFBR2 promoter induces promoter activity, whereas EWSR1-FLI1 leads to suppression of TGFBR2 promoter activity and FLI1-induced promoter activity. Introduction of EWSR1-FLI1 into cells lacking the EWSR1-FLI1 fusion suppresses TGF-β RII expression, whereas antisense to EWSR1-FLI1 in ES cell lines positive for this gene fusion restores TGF-β RII expression. Furthermore, introduction of normal TGF-β RII into ES cell lines restores TGF-β sensitivity and blocks tumorigenicity. Our results implicate TGF-β RII as a direct target of EWS-FLI1.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Ewing tumour cells with the EWSR1-FLI1 fusion are less sensitive to TGF-β.
Figure 2: Regulation of the TGFBR2 promoter by EWS-FLI1 and FLI1, and electrophoretic mobility shift assay of in vitro-translated EWS-FLI1 or FLI1.
Figure 3: Expression of TGFBR2 mRNA in CCL-64 mink epithelial cells, CHP-134B cells infected with MFG-EWS-FLI1 and in TC-32 cells infected with MFG-asEWS-FLI1.
Figure 4: Overexpression of TGF-β RII in TC32 cells restores TGF-β sensitivity and inhibits tumorigenicity.
Figure 5: Expression of TGF-β RII in primary Ewing sarcomas and neuroblastoma.
Figure 6: Features of xenograft tumours originated from TC32-CAT and TC32-TGF-β RII cells.

Similar content being viewed by others

References

  1. Rabbitts, T.H. Chromosomal translocations in human cancer. Nature 372, 143–149 (1994).

    Article  CAS  Google Scholar 

  2. Delattre, O. et al. Gene fusion with an ETS DNA binding domain caused by chromosome translocation in human cancers. Nature 359, 162–165 (1992).

    Article  CAS  Google Scholar 

  3. Sorensen, P.H.B. et al. A second Ewing's sarcoma translocation, t(21;22), fuses the EWS gene to another ETS-family transcription factor, ERG. Nature Genet. 6, 146–151 (1994).

    Article  CAS  Google Scholar 

  4. Jeon, I.-S. et al. A variant Ewing's sarcoma translocation (7;22) fuses the EWS gene to the ETS gene ETV1. Oncogene 10, 1229–1234 (1995).

    CAS  PubMed  Google Scholar 

  5. May, W. et al. Ewing sarcoma 11;22 translocation produces a chimeric transcription factor that requires the DNA-binding domain encoded by FLI1 for transformation. Proc. Natl Acad. Sci. USA 90, 5752–5756 (1993).

    Article  CAS  Google Scholar 

  6. Bailly, R.-A. et al. DNA-binding and transcriptional activation properties of the EWS-FLI-1 fusion protein resulting from the t(11;22) translocation in Ewing sarcoma. Mol. Cell. Biol. 14, 3230–3241 (1994).

    Article  CAS  Google Scholar 

  7. Braun, B.S., Frieden, R., Lessnick, S.L., May, W.A. & Denny, C.T. Identification of target genes for the Ewing's sarcoma EWS-FLI1 fusion protein by representational difference analysis. Mol. Cell. Biol. 15, 4623–4630 (1995).

    Article  CAS  Google Scholar 

  8. May, W.A. et al. EWS/FLI1-induced manic fringe renders NIH-3T3 cells tumourigenic. Nature Genet. 17, 495–497 (1997).

    Article  CAS  Google Scholar 

  9. Choi, S.-G. et al. A novel ets-related transcription factor, ERT/ESX/ESE-1, regulates expression of the transforming growth factor-β type II receptor. J. Biol. Chem. 273, 110–117 (1998).

    Article  CAS  Google Scholar 

  10. Chang, C.-H. et al. ESX: a structurally unique Ets overexpressed early during human breast tumourigenesis. Oncogene 14, 1617–1622 (1997).

    Article  CAS  Google Scholar 

  11. Oettgen, P. et al. Isolation and characterization of a novel epithelial specific transcription factor, ESE-1, a member of the ets family. Mol. Cell. Biol. 17, 4419–4433 (1997).

    Article  CAS  Google Scholar 

  12. Tymms, M.J. et al. A novel epithelial-expressed ETS gene, ELF3: human and murine cDNA sequences, murine genomic organization, human mapping to 1q32.2 and expression in tissues and cancer. Oncogene 15, 2449–2462 (1997).

    Article  CAS  Google Scholar 

  13. Andreoli, J.M. et al. The expression of a novel, epithelium-specific ets transcription factor is restricted to the most differentiated layers in the epidermis. Nucleic Acids Res. 25, 4287–4295 (1997).

    Article  CAS  Google Scholar 

  14. Bae, H.W. et al. Characterization of the promoter region of the human transforming growth factor-β type II receptor gene. J. Biol. Chem. 270, 29460–29468 (1995).

    Article  CAS  Google Scholar 

  15. Zawel, L. et al. Human Smad3 and Smad4 are sequence-specific transcription activators. Mol. Cell 1, 611–617 (1998).

    Article  CAS  Google Scholar 

  16. De Alava, E. et al. EWS/FLI1 fusion transcript structure is an independent determinant of progrosis in Ewing's sarcoma. J. Clin. Oncol 16, 1248–1255 (1998).

    Article  CAS  Google Scholar 

  17. Thompson, A.D. et al. EAT-2 is a novel SH2 domain containing protein that is up regulated by Ewing's sarcoma EWS/FLI1 fusion gene. Oncogene 15, 2649–2658 (1996).

    Google Scholar 

  18. Arvand, A. et al. EWS/FLI1 up regulates mE2-C, a cyclin-selective ubiquitin conjugating enzyme involved in cyclin B destruction. Oncogene 17, 2039–2045.

  19. Chang, J. et al. Expression of TGF-β type II receptor reduces tumourigenicity in human gastric cancer cells. Cancer Res. 57, 1–4 (1998).

    Google Scholar 

  20. Derynck, R. et al. Human transforming growth factor-β cDNA sequence and expression in tumour cell lines. Nature 316, 701–705 (1985).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank D. Wilson and S.J. Baker for FLI1 and EWSR1-FLI1 expression vectors; S.E. Kern for the SBE4-Luc construct; and A.B. Roberts, T. Parks and J. Letterio for discussion and critical review of the manuscript.

Author information

Authors and Affiliations

  1. Laboratory of Cell Regulation and Carcinogenesis, DBS, National Cancer Institute, Bethesda, 20892-5055, Maryland, USA

    Ki-Baik Hahm, Keuna Cho, Cecile Lee, Young-Hyuck Im, Jay Chang, Shin-Geon Choi & Seong-Jin Kim

  2. Pediatric Oncology Branch, DCS, National Cancer Institute, Bethesda, 20892-5055, Maryland, USA

    Carol J. Thiele

  3. Department of Pathology, British Columbia's Children's Hospital, Vancouver, V6H 3V4, British Columbia, Canada

    Poul H.B. Sorensen

Authors
  1. Ki-Baik Hahm
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Keuna Cho
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Cecile Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Young-Hyuck Im
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jay Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shin-Geon Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Poul H.B. Sorensen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Carol J. Thiele
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Seong-Jin Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Seong-Jin Kim.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hahm, KB., Cho, K., Lee, C. et al. Repression of the gene encoding the TGF-β type II receptor is a major target of the EWS-FLI1 oncoprotein. Nat Genet 23, 222–227 (1999). https://doi.org/10.1038/13854

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/13854

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing