Department of Biological Sciences, University of South Carolina, Columbia, SC, USA.
Department of Biological Sciences, University of South Carolina, Columbia, SC, USA.
Gene. 2014 Mar 15;538(1):138-49. doi: 10.1016/j.gene.2013.12.046. Epub 2014 Jan 10.
The p53 protein is an important tumor suppressor that regulates many cellular processes including maintenance of the cell cycle and apoptosis. The p53 protein maintains stability of the genome through the induction of several intracellular and extracellular factors in response to DNA damage, which in turn, stabilizes the protein, allowing it to undergo its proper mechanisms of action. We recently reported that the p53 tumor suppressor gene has a bidirectional gene partner, Wrap53β. This discovery prompted the development of a bidirectional expression vector system (pLucRLuc) that is capable of measuring the output of transcripts mediated by bidirectional promoters. We have begun to study the nature of the p53/Wrap53β promoters. Here, we have continued these studies by incorporating mutations within the regulatory regions of the p53/Wrap53β bidirectional gene pair to study the effect(s) these have on the two promoters. Deletions and point mutations were created within the two promoters and their activity was examined in the pLucRLuc system. Our results demonstrated that each of the deleted sequences within the murine p53/Wrap53β promoters reduced the activity of each of the promoters. Co-transfections with the pLucRLuc:p53/Wrap53β bidirectional expression vector and known p53 transcriptional regulators were also performed. Here, we demonstrate that p53's bidirectional gene partner, Wrap53β, can also be regulated by many of the same transcription factors.
p53 蛋白是一种重要的肿瘤抑制因子,它调节许多细胞过程,包括细胞周期的维持和细胞凋亡。p53 蛋白通过诱导几种细胞内和细胞外因子来维持基因组的稳定性,这些因子反过来又稳定了蛋白质,使其能够进行适当的作用机制。我们最近报道称,p53 肿瘤抑制基因有一个双向基因伙伴 Wrap53β。这一发现促使我们开发了一种双向表达载体系统(pLucRLuc),该系统能够测量由双向启动子介导的转录本的输出。我们已经开始研究 p53/Wrap53β 启动子的性质。在这里,我们通过在 p53/Wrap53β 双向基因对的调节区域内引入突变,继续研究这些突变对两个启动子的影响。在两个启动子内创建了缺失和点突变,并在 pLucRLuc 系统中检查了它们的活性。我们的结果表明,在小鼠 p53/Wrap53β 启动子内的每个缺失序列都降低了每个启动子的活性。还进行了 pLucRLuc:p53/Wrap53β 双向表达载体与已知 p53 转录调节剂的共转染。在这里,我们证明 p53 的双向基因伙伴 Wrap53β 也可以被许多相同的转录因子调节。