Huet X, Rech J, Plet A, Vié A, Blanchard J M
Institut de Génétique Moléculaire de Montpellier, CNRS UMR 5535, France.
Mol Cell Biol. 1996 Jul;16(7):3789-98. doi: 10.1128/MCB.16.7.3789.
Transcription of the gene coding for cyclin A, a protein required for S-phase transit, is cell cycle regulated and is restricted to proliferating cells. To further explore transcriptional regulation linked to cell division cycle control, a genomic clone containing 5' flanking sequences of the murine cyclin A gene was isolated. When it was fused to a luciferase reporter gene, it was shown to function as a proliferation-regulated promoter in NIH 3T3 cells. Transcription of the mouse cyclin A gene is negatively regulated by arrest of cell proliferation. A mutation of a GC-rich sequence conserved between mice and humans is sufficient to relieve transcriptional repression, resulting in a promoter with constitutively high activity. In agreement with this result, in vivo footprinting reveals a protection of the cell cycle-responsive element in G0/early G1 cells which is not observed at later stages of the cell cycle. Moreover, the footprint is present in dimethyl sulfoxide-induced differentiating and not in proliferating Friend erythroleukemia cells. Conversely, two other sites, which in vitro bind ATF-1 and NF-Y, respectively, are constitutively occupied throughout cell cycle progression.
细胞周期蛋白A编码基因的转录在细胞周期中受到调控,且仅限于增殖细胞,细胞周期蛋白A是S期过渡所需的一种蛋白质。为了进一步探索与细胞分裂周期控制相关的转录调控,分离出了一个包含小鼠细胞周期蛋白A基因5'侧翼序列的基因组克隆。当它与荧光素酶报告基因融合时,在NIH 3T3细胞中显示出作为增殖调控启动子的功能。小鼠细胞周期蛋白A基因的转录受到细胞增殖停滞的负调控。小鼠和人类之间保守的富含GC序列的突变足以解除转录抑制,从而产生一个具有组成型高活性的启动子。与这一结果一致,体内足迹分析显示在G0/早期G1细胞中细胞周期反应元件受到保护,而在细胞周期后期未观察到这种保护。此外,在二甲基亚砜诱导分化的弗氏红白血病细胞中存在足迹,而在增殖细胞中不存在。相反,另外两个分别在体外与ATF-1和NF-Y结合的位点在整个细胞周期进程中都被组成型占据。