Nucleic Acids Research, Vol 24, Issue 15 2950-2958, Copyright © 1996 by Oxford University Press
J Bernues, P Carrera and F Azorin
The binding of TBP (TFIID) to the TATA box has been considered to direct
promoter recognition and pre-initiation complex formation because it is the
first event leading to basal transcription by RNA polymerase II. Here, we
analyse the binding of yeast TBP to a consensus TATAAA box and two point
mutations, TAAAAA (inactive) and TATATA (active). Despite the fact that the
TAAAAA sequence does not support transcription in vitro, yeast TBP binds
the three sequences showing, in this sense, only a limited sequence
specificity. However, the TBP- TAAAAA complex cannot be recognised by other
basal transcription factors, in particular by TFIIB. DNase I footprinting
patterns of the TBP-TAAAAA complex are different from those observed in
functional TBP- TATA box complexes, indicating that, most likely, it is a
different spatial arrangement of the TBP-DNA complex that prevents
formation of the TFIIB-TBP-TAAAAA complex, also seriously impairing entry
of TFIIA to the complex. DNA deformability of the A/T-rich sequences
appears to be an important determinant in the formation of a productive
TBP-TATA complex. These results indicate that the transcriptional
competence of A/T-rich sequences is determined not only by TBP binding, but
also by the ability of other basal transcription factors to recognise the
preformed TBP-DNA complexes.
ARTICLES
TBP binds the transcriptionally inactive TA5 sequence but the resulting complex is not efficiently recognised by TFIIB and TFIIA
Dept. Biologie Molecular i Cellular, Centre d'Investigacio i Desenvolupament CSIC, Barcelona, Spain.
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