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J. Biol. Chem., Vol. 269, Issue 42, 26575-26583, 10, 1994
T Aso, JW Conaway and RC Conaway
Efforts to understand the impact of core promoter architecture on the
mechanism of transcription initiation by RNA polymerase II have been
hampered by lack of well defined, reconstituted transcription systems
responsive both to efficiently transcribed consensus and near consensus
TATA box-containing promoters and to considerably weaker TATA-less
promoters. In this report, we investigate the influence of core promoter
structure on the mechanism of assembly of the RNA polymerase II
preinitiation complex using a highly purified, holoTFIID-dependent
transcription system that permits sensitive measurement of transcription
initiation from a wide variety of TATA and TATA-less promoters in the
absence of transactivators. A direct comparison of the requirements for
formation of stable preinitiation intermediates at these promoters led to
the discovery that, whereas holoTFIID binds avidly to the consensus TATA-
and strong initiator-containing adenovirus major late (AdML) promoter to
form the first stable intermediate on the pathway leading to formation of
the complete preinitiation complex, it binds poorly not only to TATA-less
promoters but also to promoters with consensus or near consensus TATA
elements. With the exception of the AdML promoter, formation of stable
preinitiation intermediates at each of the promoters tested was found to be
strongly dependent on RNA polymerase II, holoTFIID, and TFIIB and was
stimulated by TFIIF. Based on these observations, we suggest that RNA
polymerase II assembles with many TATA and TATA-less promoters by a common
pathway.
Role of core promoter structure in assembly of the RNA polymerase II preinitiation complex. A common pathway for formation of preinitiation intermediates at many TATA and TATA-less promoters
Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, Oklahoma City 73104.
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