THE REQUIREMENT FOR THE BASAL TRANSCRIPTION FACTOR IIE IS DETERMINED BY THE HELICAL STABILITY OF PROMOTER DNA
THE REQUIREMENT FOR THE BASAL TRANSCRIPTION FACTOR IIE IS DETERMINED BY THE HELICAL STABILITY OF PROMOTER DNA
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DOI：
10.1002/j.1460-2075.1995.tb07059.x
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发表时间：
1995-02-15
期刊：
影响因子：
11.4
通讯作者：
TIMMERS, HTM
中科院分区：
文献类型：
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作者：
HOLSTEGE, FCP;TANTIN, D;TIMMERS, HTM
研究了基础转录因子TFIIE在用纯化蛋白质重建的RNA聚合酶II转录反应中的作用。使用负超螺旋模板，这规避了TFIIH的要求，我们观察到，从腺病毒主要晚期（ML）核心启动子的转录比从腺病毒E4（E4）或小鼠乳腺肿瘤病毒（MMTV）启动子的转录更依赖于TFIIE。对于所有三种促进剂，反应混合物的离子强度的增加导致对TFIIE的依赖性增加。混合ML/MMTV启动子的分析表明，从-10到+10的起始位点区域决定了这种依赖性。从具有预熔化的-8至+2区域的松弛E4模板的转录完全独立于TFIIE和TFIIH两者。我们建议，负超螺旋模板TFIIE可以促进启动子熔化。
The role of the basal transcription factor TFIIE was investigated in RNA polymerase II transcription reactions reconstituted with purified proteins. Using negatively supercoiled templates, which circumvent the requirement for TFIIH, we observed that transcription from the adenovirus major-late (ML) core promoter is more dependent on TFIIE than transcription from the adenovirus E4 (E4) or mouse mammary tumor virus (MMTV) promoters. For all three promoters, an increase in the ionic strength of the reaction mixtures led to an increased dependence on TFIIE. Analysis of hybrid ML/MMTV promoters showed that the region encompassing the start site, from -10 to +10, dictates this dependence. Transcription from a relaxed E4 template with a pre-melted -8 to +2 region was completely independent of both TFIIE and TFIIH. We propose that on negatively supercoiled templates TFIIE can facilitate promoter melting.