DNA binding domain-independent pathways are involved in EWS/FLI1-mediated oncogenesis.
Specific chromosomal translocations involving the ews gene and one of five members of the ets family of transcription factors create ews/ets fusion genes that are found in approximately 85% of Ewing's family of tumors. ews/ets fusion genes consistently maintain an intact and functional ets DNA binding domain (DBD) in all of these cases. We demonstrate here, however, that EWS/FLI1, the most prevalent EWS/ETS fusion, activates oncogenic pathways independent of its DBD. In in vivo tumor assays, EWS/FLI1 molecules with either point mutations or a large deletion in the ets DBD retain the ability to accelerate tumors in NIH 3T3 cells, whereas they lose the ability to bind DNA in vitro. Additionally, whereas inhibition of DBD functions of EWS/FLI1 with a dominant negative form of FLI1 is sufficient to inhibit anchorage-independent growth in NIH 3T3 cells, it is ineffective in inhibiting tumor growth in SCID mice. Usage of this dominant negative construct in a Ewing's tumor cell line, however, does reduce the rate of tumor formation, supporting the need for a functional DBD in this context. Together, these results suggest that EWS/FLI1 induces both DBD-dependent and DBD-independent oncogenic pathways.
Welford
S M
SM
Molecular Biology Institute, UCLA, Los Angeles, California 90095, USA.
Hebert
S P
SP
Deneen
B
B
Arvand
A
A
Denny
C T
CT
D016475:3T3 Cells; D000818:Animals; D001665:Binding Sites; D002471:Cell Transformation, Neoplastic; D004247:DNA / Q000378:metabolism*; D005260:Female; D051379:Mice; D016513:Mice, SCID; D009374:Neoplasms, Experimental / Q000209:etiology*; D015514:Oncogene Proteins, Fusion / Q000737:chemistry / Q000502:physiology*; D051789:Proto-Oncogene Protein c-fli-1; D034802:RNA-Binding Protein EWS; D012512:Sarcoma, Ewing / Q000209:etiology; D014157:Transcription Factors / Q000737:chemistry / Q000502:physiology*
C106031:EWS-FLI fusion protein; D015514:Oncogene Proteins, Fusion; D051789:Proto-Oncogene Protein c-fli-1; D034802:RNA-Binding Protein EWS; D014157:Transcription Factors; D004247:DNA