酿酒酵母TFIIF在转录起始位点利用过程中的功能。
Functions of Saccharomyces cerevisiae TFIIF during transcription start site utilization.
作者信息
Khaperskyy Denys A, Ammerman Michelle L, Majovski Robert C, Ponticelli Alfred S
机构信息
Department of Biochemistry, School of Medicine and Biomedical Sciences, State University of New York, Buffalo, NY 14214-3000, USA.
出版信息
Mol Cell Biol. 2008 Jun;28(11):3757-66. doi: 10.1128/MCB.02272-07. Epub 2008 Mar 24.
Abstract
Previous studies have shown that substitutions in the Tfg1 or Tfg2 subunits of Saccharomyces cerevisiae transcription factor IIF (TFIIF) can cause upstream shifts in start site utilization, resulting in initiation patterns that more closely resemble those of higher eukaryotes. In this study, we report the results from multiple biochemical assays analyzing the activities of wild-type yeast TFIIF and the TFIIF Tfg1 mutant containing the E346A substitution (Tfg1-E346A). We demonstrate that TFIIF stimulates formation of the first two phosphodiester bonds and dramatically stabilizes a short RNA-DNA hybrid in the RNA polymerase II (RNAPII) active center and, importantly, that the Tfg1-E346A substitution coordinately enhances early bond formation and the processivity of early elongation in vitro. These results are discussed within a proposed model for the role of yeast TFIIF in modulating conformational changes in the RNAPII active center during initiation and early elongation.
摘要
先前的研究表明，酿酒酵母转录因子IIF（TFIIF）的Tfg1或Tfg2亚基中的取代可导致起始位点利用的上游偏移，从而产生更类似于高等真核生物的起始模式。在本研究中，我们报告了多项生化分析的结果，这些分析检测了野生型酵母TFIIF和含有E346A取代的TFIIF Tfg1突变体（Tfg1-E346A）的活性。我们证明，TFIIF刺激前两个磷酸二酯键的形成，并显著稳定RNA聚合酶II（RNAPII）活性中心的短RNA-DNA杂交体，重要的是，Tfg1-E346A取代在体外协同增强早期键的形成和早期延伸的持续性。我们在一个关于酵母TFIIF在起始和早期延伸过程中调节RNAPII活性中心构象变化作用的模型中讨论了这些结果。
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