Imbalzano A N, Zaret K S, Kingston R E
Department of Molecular Biology, Massachusetts General Hospital, Boston 02114.
J Biol Chem. 1994 Mar 18;269(11):8280-6.
The general transcription factors (TF) IIB and TFIIA are the first factors to associate with the TATA-binding protein (TBP) during formation of a transcription initiation complex on RNA polymerase II promoters. DNase I footprint titration was used to measure the effects of TFIIB and TFIIA on binding of TBP to a consensus TATA box. Under reaction conditions optimized for TBP-DNA complex formation, the presence of TFIIB increased affinity of TBP for the TATA box by 2.5-fold, while TFIIA had no effect. When TBP binding conditions were sub-optimal, both TFIIB and TFIIA independently increased TBP affinity by approximately 10-fold. Therefore both TFIIB and TFIIA have the intrinsic ability to directly increase the affinity of TBP for the TATA box. We suggest that this property of TFIIA and TFIIB may increase the range of conditions under which high affinity TBP-DNA interactions can occur and may therefore favor the formation of the preinitiation complex.
通用转录因子IIB(TFIIB)和IIA(TFIIA)是在RNA聚合酶II启动子上形成转录起始复合物过程中首先与TATA结合蛋白(TBP)结合的因子。使用DNase I足迹滴定法来测量TFIIB和TFIIA对TBP与共有TATA框结合的影响。在针对TBP-DNA复合物形成优化的反应条件下,TFIIB的存在使TBP对TATA框的亲和力增加了2.5倍,而TFIIA没有影响。当TBP结合条件次优时,TFIIB和TFIIA都能独立地使TBP亲和力增加约10倍。因此,TFIIB和TFIIA都具有直接增加TBP对TATA框亲和力的内在能力。我们认为,TFIIA和TFIIB的这一特性可能会增加高亲和力TBP-DNA相互作用能够发生的条件范围,因此可能有利于预起始复合物的形成。