Ets2 is not required for Ras or Neu/ErbB-2 mediated cellular transformation in vitro

Ets2 is a widely expressed Ets family transcription factor which is activated by Ras signaling and has been reported to transform fibroblasts. Expression of a dominant inhibitory Ets2 construct consisting of just the Ets2 DNA binding domain (Ets2DBD), reverses Ras transformation of NIH3T3 cells and the transformed characteristics of several human tumor cells. However, the Ets2DBD may interfere with multiple Ets family members. We have now utilized cell lines with a disrupted ets2 gene to determine whether Ets2 is required to mediate oncogenic signaling. Expression of the Ets2DBD in an Ets2-deficient cell line dramatically inhibited Ets-dependent (but not AP-1-dependent) reporter gene expression, revealing that the Ets2DBD does inhibit additional Ets family members. The transformation efficiency of Ets2-deficient cell lines by oncogenic Ras or Neu/ErbB-2 was similar to that of control cells in several in vitro assays, and was not enhanced by re-expression of Ets2. Finally, overexpression of Ets2 was not sufficient to induce focus formation in NIH3T3 cells, nor to enhance transformation by oncogenic Ras. Thus, Ets2 is not an essential mediator of Ras or Neu/ErbB-2 transformation in these cells. Our results illustrate the importance of utilizing specific approaches for analyzing the function of individual members of large gene families.