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Transforming activity of EWS/FLI is not strictly dependent upon DNA-binding activity

Oncogene volume 18, pages 5592–5597 (1999)Cite this article

Abstract

In approximately 85% of Ewing sarcomas, chromosomal translocations give rise to the chimeric gene EWS/FLI, encoding the N-terminus of the RNA binding protein EWS fused to the DNA-binding domain of the ETS protein FLI-1. EWS/FLI is a stronger transcriptional activator than wild-type FLI-1, although both proteins bind to the same DNA sequences in vitro. In addition, EWS/FLI, but not FLI-1, is a transforming oncogene in NIH3T3 fibroblasts. EWS/FLI is thought to transform through its ability to deregulate the expression of target genes. We introduced several point mutations into the ETS domain of EWS/FLI that abolished DNA-binding activity. Although two of these mutations disrupted the transforming activity of EWS/FLI, one mutated protein containing a substitution of isoleucine 347 with glutamic acid (I347E) retained diminished transforming activity. In addition, EWS/FLI I347E did not activate expression of the endogenous EWS/FLI target gene manic fringe (MFNG). These studies demonstrate that a portion of the oncogenic activity of EWS/FLI is independent of FLI DNA-binding activity.

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Acknowledgements

We thank Frank Rauscher, The Wistar Institute, for pCB6+, Charles Sawyers, UCLA Medical Center, for pSRα(ΔHindIII)-tk-Neo retroviral vector, David Baltimore, California Institute of Technology, for 293T cells, Clare Sample, St Jude Children's Research Hospital, for PU.1 cDNA, and Timothy J Triche, Children's Hospital, USC, for TC252 cells. We also thank Tom Curran, Peter J McKinnon and Stephen W White for helpful discussions. This work was supported in part by NIH grants PO1-CA-71907 (SJ Baker and MF Roussel), the Cancer Center Support CORE grant P30 CA21765 and by the American Lebanese Syrian Associated Charities (ALSAC).

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Authors and Affiliations

  1. Department of Developmental Neurobiology, St Jude Children's Research Hospital, Memphis, TN 38105, Tennessee, USA

    S Jaishankar, J Zhang & S J Baker

  2. Department of Tumor Cell Biology, St Jude Children's Research Hospital, Memphis, TN 38105, Tennessee, USA

    M F Roussel

  3. Department of Pathology, University of Tennessee, Memphis, TN 38163, Tennessee, USA

    S J Baker

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  1. S Jaishankar
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Jaishankar, S., Zhang, J., Roussel, M. et al. Transforming activity of EWS/FLI is not strictly dependent upon DNA-binding activity. Oncogene 18, 5592–5597 (1999). https://doi.org/10.1038/sj.onc.1202940

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