Klug J, Beato M
Philipps-Universität Marburg, Institut für Molekularbiologie und Tumorforschung, Germany.
Mol Cell Biol. 1996 Nov;16(11):6398-407. doi: 10.1128/MCB.16.11.6398.
The gene for rabbit uteroglobin codes for a small calcium-, steroid-, and biphenyl metabolite-binding homodimeric protein which is expressed in a variety of epithelial cell types such as Clara cells (lung) and the glandular and luminal cells of the endometrium. One important region mediating its efficient transcription in a human endometrium-derived cell line, Ishikawa, is centered around a noncanonical TATA box. Two factors, TATA core factor (TCF), expressed in cell lines derived from uteroglobin-expressing tissues, and the ubiquitously expressed TATA palindrome factor, bind to the DNA major groove at two adjacent sites within this region. Here, we report the identification of the TATA palindrome factor as the transcription/initiation factor YY1 by microsequencing of the biochemically purified factor from HeLa cells. The binding site for YY1 within the uteroglobin gene is unique in its sequence and its location overlapping a weak TATA box (TACA). Binding of YY1 was required for efficient transcription in TCF-positive Ishikawa cells, which responded only weakly to a change of TACA to TATA, although in vitro binding affinity for the TATA-box-binding protein increased by 1 order of magnitude. In contrast, in CV-1 cells, lacking TCF, binding of YY1 was not required for transcription in the context of a wild-type TACA box, whereas a change from TACA to TATA led to significantly increased reporter gene expression. DNA binding data exclude a role of YY1 in stabilizing the interaction of the TATA-box-binding protein with the uteroglobin promoter. We conclude that cell lines derived from uteroglobin-expressing tissues overcome the weak TATA box with the help of auxiliary factors, one of them being YY1.
兔子宫珠蛋白基因编码一种小的同型二聚体蛋白,该蛋白可结合钙、类固醇和联苯代谢物,在多种上皮细胞类型中表达,如克拉拉细胞(肺)以及子宫内膜的腺细胞和腔细胞。在源自人子宫内膜的细胞系 Ishikawa 中,介导其高效转录的一个重要区域围绕一个非典型 TATA 框。两种因子,即表达于源自子宫珠蛋白表达组织的细胞系中的 TATA 核心因子(TCF)和普遍表达的 TATA 回文因子,在该区域内的两个相邻位点与 DNA 大沟结合。在此,我们通过对从 HeLa 细胞中生化纯化的因子进行微量测序,报告了 TATA 回文因子被鉴定为转录/起始因子 YY1。子宫珠蛋白基因内 YY1 的结合位点在序列和位置上都很独特,它与一个弱 TATA 框(TACA)重叠。在 TCF 阳性的 Ishikawa 细胞中,YY1 的结合是高效转录所必需的,尽管 TACA 变为 TATA 时体外与 TATA 框结合蛋白的结合亲和力增加了 1 个数量级,但这些细胞对这种变化反应较弱。相反,在缺乏 TCF 的 CV - 1 细胞中,在野生型 TACA 框的情况下,YY1 的结合对于转录不是必需的,而从 TACA 变为 TATA 则导致报告基因表达显著增加。DNA 结合数据排除了 YY1 在稳定 TATA 框结合蛋白与子宫珠蛋白启动子相互作用中的作用。我们得出结论,源自子宫珠蛋白表达组织的细胞系借助辅助因子克服了弱 TATA 框,其中之一就是 YY1。