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J. Biol. Chem., Vol. 275, Issue 45, 35006-35012, November 10, 2000
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From the The TATA-binding protein (TBP) in the TFIID
complex binds specifically to the TATA-box to initiate the stepwise
assembly of the preinitiation complex (PIC) for RNA polymerase II
transcription. Transcriptional activators and repressors compete with
general transcription factors at each step to influence the course of the assembly. To investigate this process, the TBP·TATA complex was
titrated with HMG-1 and the interaction monitored by electrophoretic mobility shift assays. The titration produced a ternary
HMG-1·TBP·TATA complex, which exhibits increased mobility relative
to the TBP·TATA complex. The addition of increasing levels of TFIIB
to this complex results in the formation of the TFIIB·TBP·TATA
complex. However, in the reverse titration, with very high mole
ratios of HMG-1 present, TFIIB is not dissociated off and a
complex is formed that contains all factors. The simultaneous addition
of E1A to a mixture of TBP and TATA; or HMG-1, TBP, and TATA; or TFIIB, TBP, and TATA inhibits complex formation. On the other hand, E1A added
to the pre-established complexes shows a significantly reduced capability to disrupt the complex. In add-back experiments with all
complexes, increased levels of TBP re-established the complexes, indicating that the primary target for E1A in all complexes is TBP.
Influence of HMG-1 and Adenovirus Oncoprotein E1A on Early Stages
of Transcriptional Preinitiation Complex Assembly*
,, and¶
Department of Chemistry, Bowling Green State
University, Bowling Green, Ohio 43403 and the
§ Department of Chemistry, Ohio Northern University, Ada,
Ohio 45810
*
This work was supported by Grant R15 from the National
Institutes of Health (to W. M. S.) and by grants from the Ohio Cancer Research Associates and the American Cancer Society, Ohio
Division.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.
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