Mutation in the DNA-binding domain of the EWS-Oct-4 oncogene results in dominant negative activity that interferes with EWS-Oct-4-mediated transactivation

Int J Cancer. 2009 May 15;124(10):2312-22. doi: 10.1002/ijc.24228.


The EWS-Oct-4 protein is a chimeric molecule in which the amino terminal domain (NTD) of the EWS becomes fused to the carboxy terminal domain (CTD) of the Oct-4 transcription factor. It was identified in human bone and soft-tissue tumors associated with t(6;22)(p21;q12). Using in vitro and in vivo systems, we found that the EWS-Oct-4 protein self-associates. The major domains required for self-association mapped to the EWS NTD (amino acids 70-163) and the POU DNA-binding domain. EWS-Oct-4 protein also associated with EWS-Oct-4 (V351P), which contains a mutation in the POU DNA-binding domain. Using electrophoretic mobility shift assays, we found that the EWS-Oct-4 (V351P) mutant interfered with wild-type EWS-Oct-4 DNA-binding activity. In addition, we found that EWS-Oct-4-mediated transcriptional activation was inhibited by EWS-Oct-4 (V351P) protein in vivo. Thus, this mutation in the POU DNA-binding domain results in a dominant negative protein. These findings suggest that the biological functions of the EWS-Oct-4 oncogene can be modulated by the dominant negative mutant EWS-Oct-4 (V351P).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Chromosomes, Human, Pair 22
  • Chromosomes, Human, Pair 6
  • DNA / metabolism*
  • DNA Primers
  • Electrophoretic Mobility Shift Assay
  • Genes, Dominant*
  • Humans
  • Mutagenesis, Site-Directed
  • Octamer Transcription Factor-3 / genetics*
  • Octamer Transcription Factor-3 / physiology
  • Oncogenes*
  • RNA-Binding Protein EWS / genetics*
  • RNA-Binding Protein EWS / physiology
  • Transcriptional Activation / physiology*


  • DNA Primers
  • Octamer Transcription Factor-3
  • RNA-Binding Protein EWS
  • DNA