Open AccessEWS, but Not EWS-FLI-1, Is Associated with Both TFIID and RNA Polymerase II: Interactions between Two Members of the TET Family, EWS and hTAFII68, and Subunits of TFIID and RNA Polymerase II Complexes
Author(s) -
Anne Bertolotti,
Thomas Melot,
Joël Acker,
Marc Vigneron,
Olivier Delattre,
Làszlò Tora
Publication year - 1998
Publication title -
molecular and cellular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.14
H-Index - 327
eISSN - 1067-8824
pISSN - 0270-7306
DOI - 10.1128/mcb.18.3.1489
Subject(s) - taf1 , transcription factor ii d , biology , transcription factor ii a , rna polymerase ii , genetics , microbiology and biotechnology , tata box binding protein , general transcription factor , transcription factor , gene , rna polymerase , dna binding protein , rna , gene expression , promoter
The t(11;22) chromosomal translocation specifically linked to Ewing sarcoma and primitive neuroectodermal tumor results in a chimeric molecule fusing the amino-terminus-encoding region of theEWS gene to the carboxyl-terminal DNA-binding domain encoded by theFLI-1 gene. As the function of the protein encoded by theEWS gene remains unknown, we investigated the putative role of EWS in RNA polymerase II (Pol II) transcription by comparing its activity with that of its structural homolog, hTAFII 68. We demonstrate that a portion of EWS is able to associate with the basal transcription factor TFIID, which is composed of the TATA-binding protein (TBP) and TBP-associated factors (TAFII s). In vitro binding studies revealed that both EWS and hTAFII 68 interact with the same TFIID subunits, suggesting that the presence of EWS and that of hTAFII 68 in the same TFIID complex may be mutually exclusive. Moreover, EWS is not exclusively associated with TFIID but, similarly to hTAFII 68, is also associated with the Pol II complex. The subunits of Pol II that interact with EWS and hTAFII 68 have been identified, confirming the association with the polymerase. In contrast to EWS, the tumorigenic EWS–FLI-1 fusion protein is not associated with either TFIID or Pol II in Ewing cell nuclear extracts. These observations suggest that EWS and EWS–FLI-1 may play different roles in Pol II transcription.