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Mol Cell Biol, March 1998, p. 1489-1497, Vol. 18, No. 3
Institut de Génétique et de
Biologie Moléculaire et Cellulaire,
Received 4 September 1997/Returned for modification 14 October
1997/Accepted 24 November 1997
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 the
EWS gene to the carboxyl-terminal DNA-binding domain
encoded by the FLI-1 gene. As the function of the protein
encoded by the EWS 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,
hTAFII68. 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
(TAFIIs). In vitro binding studies revealed that both EWS
and hTAFII68 interact with the same TFIID subunits,
suggesting that the presence of EWS and that of hTAFII68 in
the same TFIID complex may be mutually exclusive. Moreover, EWS is not
exclusively associated with TFIID but, similarly to hTAFII68, is also associated with the Pol II complex. The
subunits of Pol II that interact with EWS and hTAFII68 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.
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
EWS, 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
*
Corresponding author. Mailing address: Institut de
Génétique et de Biologie Moléculaire et Cellulaire,
CNRS/INSERM/ULP, BP 163, F-67404 ILLKIRCH Cedex, C.U. de Strasbourg,
France. Phone: (33) 3 88 65 34 44. Fax: (33) 3 88 65 32 01. E-mail:
laszlo{at}titus.u-strasbg.fr.
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