Martin J, Halenbeck R, Kaufmann J
Chiron Corporation, Chiron Technologies, Emeryville, California 94608, USA.
Mol Cell Biol. 1999 Aug;19(8):5548-56. doi: 10.1128/MCB.19.8.5548.
Here we present evidence that CIF150 (hTAF(II)150), the human homolog of Drosophila TAF(II)150, plays an important and selective role in establishing gene expression patterns necessary for progression through the cell cycle. Gel filtration experiments demonstrate that CIF150 (hTAF(II)150) seems to be less tightly associated with human transcription factor IID than hTAF(II)130 is associated with hTAF(II)250. The transient functional knockout of CIF150 (hTAF(II)150) protein led to cell cycle arrest at the G(2)/M transition in mammalian cell lines. PCR display analysis with the RNA derived from CIF150-depleted cells indicated that CIF150 (hTAF(II)150) is required for the transcription of only a subset of RNA polymerase II genes. CIF150 (hTAF(II)150) directly stimulated cyclin B1 and cyclin A transcription in cotransfection assays and in vitro assays, suggesting that the expression of these genes is dependent on CIF150 (hTAF(II)150) function. We defined a CIF150 (hTAF(II)150) consensus binding site and demonstrated that a CIF150-responsive cis element is present in the cyclin B1 core promoter. These results suggest that one function of CIF150 (hTAF(II)150) is to select specific RNA polymerase II core promoter elements involved in cell cycle progression.
在此,我们提供证据表明,果蝇TAF(II)150的人类同源物CIF150(hTAF(II)150)在建立细胞周期进程所需的基因表达模式中发挥重要且具有选择性的作用。凝胶过滤实验表明,与hTAF(II)130和hTAF(II)250的结合相比,CIF150(hTAF(II)150)似乎与人类转录因子IID的结合不那么紧密。CIF150(hTAF(II)150)蛋白的瞬时功能敲除导致哺乳动物细胞系在G(2)/M期转换时出现细胞周期停滞。对来自CIF150缺失细胞的RNA进行PCR显示分析表明,CIF150(hTAF(II)150)仅对RNA聚合酶II基因的一个子集的转录是必需 的。在共转染实验和体外实验中,CIF150(hTAF(II)150)直接刺激细胞周期蛋白B1和细胞周期蛋白A的转录,这表明这些基因的表达依赖于CIF150(hTAF(II)150)的功能。我们定义了一个CIF150(hTAF(II)150)共有结合位点,并证明在细胞周期蛋白B1核心启动子中存在一个CIF150反应性顺式元件。这些结果表明,CIF150(hTAF(II)150)的一个功能是选择参与细胞周期进程的特定RNA聚合酶II核心启动子元件。