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J. Biol. Chem., Vol. 278, Issue 13, 11495-11501, March 28, 2003
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From the Laboratory of Gene Transcription, Institut de Recherches
Cliniques de Montréal, Montréal, Quebec H2W 1R7,
Canada
Site-specific protein-DNA photo-cross-linking was
used to show that, when bound to its cognate site at various distances
upstream of the TATA element, the chimeric transcriptional activator
GAL4-VP16 can physically interact with a TATA box-binding protein
(TBP)- transcription factor IIA (TFIIA)-TFIIB complex assembled on the TATA element. This result implies DNA bending and looping of promoter DNA as a result of the physical interaction between GAL4-VP16 and an
interface of the TBP-TFIIA-TFIIB complex. This protein-protein interaction on promoter DNA minimally requires the presence of one GAL4
binding site and the formation of a quaternary complex containing TBP,
TFIIB, and TFIIA on the TATA element. Notably, the topology of the
TBP-TFIIA-TFIIB-promoter complex is not altered significantly by the
interaction with DNA-bound activators. We also show that the ability of
GAL4-VP16 to activate transcription through a single GAL4 binding site
varies according to its precise location and orientation relative to
the TATA element and that it can approach the efficiency obtained with
multiple binding sites. Taken together, our results indicate
that the spatial positioning of the DNA-bound activation domain is
important for efficient activation, possibly by maximizing its
interactions with the transcriptional machinery including the
TBP-TFIIA-TFIIB-promoter quaternary complex.
Interactions of a DNA-bound Transcriptional Activator with the
TBP-TFIIA-TFIIB-Promoter Quaternary Complex*
*
This work was supported by a grant from the Canadian
Institutes of Health Research.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Senior scholar from the Fonds de recherche en santé du
Québec. To whom correspondence should be addressed: Laboratory of Gene Transcription, Institut de Recherches Cliniques de Montréal, 110 Ave. des Pins Ouest, Montréal, Québec H2W 1R7, Canada.
Tel.: 514-987-5662; Fax: 514-987-5663; E-mail:
coulomb@ircm.qc.ca.
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