Takada S, Lis J T, Zhou S, Tjian R
Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, Berkeley, 94720, USA.
Cell. 2000 May 26;101(5):459-69. doi: 10.1016/s0092-8674(00)80857-0.
It has been generally accepted that the TATA binding protein (TBP) is a universal mediator of transcription by RNA polymerase I, II, and III. Here we report that the TBP-related factor TRF1 rather than TBP is responsible for RNA polymerase III transcription in Drosophila. Immunoprecipitation and in vitro transcription assays using immunodepleted extracts supplemented with recombinant proteins reveals that a TRF1:BRF complex is required to reconstitute transcription of tRNA, 5S and U6 RNA genes. In vivo, the majority of TRF1 is complexed with BRF and these two proteins colocalize at many polytene chromosome sites containing RNA pol III genes. These data suggest that in Drosophila, TRF1 rather than TBP forms a complex with BRF that plays a major role in RNA pol III transcription.
人们普遍认为,TATA 结合蛋白(TBP)是 RNA 聚合酶 I、II 和 III 转录的通用介导因子。在此我们报告,在果蝇中,与 TBP 相关的因子 TRF1 而非 TBP 负责 RNA 聚合酶 III 的转录。使用补充了重组蛋白的免疫耗尽提取物进行的免疫沉淀和体外转录分析表明,重建 tRNA、5S 和 U6 RNA 基因的转录需要 TRF1:BRF 复合物。在体内,大多数 TRF1 与 BRF 形成复合物,并且这两种蛋白在许多含有 RNA 聚合酶 III 基因的多线染色体位点共定位。这些数据表明,在果蝇中,TRF1 而非 TBP 与 BRF 形成复合物,该复合物在 RNA 聚合酶 III 的转录中起主要作用。