Maxon M E, Goodrich J A, Tjian R
Howard Hughes Medical Institute, Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Genes Dev. 1994 Mar 1;8(5):515-24. doi: 10.1101/gad.8.5.515.
The basal factor TFIIE is an important component of the RNA polymerase II transcription machinery. In our efforts to determine the role of TFIIE in the transcription process, we have begun to define the interactions between TFIIE and other basal transcription factors. Here we report that TFIIE binds selectively to the nonphosphorylated form of RNA polymerase II (IIa) and that this interaction is mediated by the 56-kD subunit of TFIIE. Additional binding studies reveal that TFII can interact with TBP as well as TFIID and that this interaction is mediated primarily via the 56-kD subunit. Our studies indicate that TFIIE also interacts with both subunits of TFIIF and with TFIIH, a multisubunit basal factor reported to catalyze RNA polymerase II CTD phosphorylation. Protein affinity assays demonstrate that TFIIE binds directly to ERCC-3, a DNA repair protein associated with TFIIH. More importantly, TFIIE affinity resin can selectively isolate transcriptionally competent TFIIH from a partially purified preparation and thereby may recruit TFIIH to the transcription complex in vivo. These multiple interactions between TFIIE, Pol II and TFIIH support a model in which TFIIE plays a role in promoter clearance as well as in the recruitment of proteins required for transcription-coupled DNA repair.
基础因子TFIIE是RNA聚合酶II转录机制的重要组成部分。在我们确定TFIIE在转录过程中作用的研究中,我们已开始明确TFIIE与其他基础转录因子之间的相互作用。在此我们报告,TFIIE选择性地结合RNA聚合酶II的非磷酸化形式(IIa),并且这种相互作用由TFIIE的56-kD亚基介导。额外的结合研究表明,TFIIE能与TBP以及TFIID相互作用,且这种相互作用主要通过56-kD亚基介导。我们的研究表明,TFIIE还与TFIIF的两个亚基以及TFIIH相互作用,TFIIH是一个据报道可催化RNA聚合酶II CTD磷酸化的多亚基基础因子。蛋白质亲和分析表明,TFIIE直接结合ERCC-3,ERCC-3是一种与TFIIH相关的DNA修复蛋白。更重要的是,TFIIE亲和树脂可从部分纯化的制剂中选择性地分离出具有转录活性的TFIIH,因此可能在体内将TFIIH招募到转录复合物中。TFIIE、Pol II和TFIIH之间的这些多重相互作用支持了一种模型,即TFIIE在启动子清除以及招募转录偶联DNA修复所需蛋白质方面发挥作用。