Sepelakova Janka, Takacova Martina, Pastorekova Silvia, Kopacek Juraj
Centre of Molecular Medicine, Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, Bratislava 845 05, Slovakia.
Biochim Biophys Acta. 2005 Dec 20;1731(3):209-14. doi: 10.1016/j.bbaexp.2005.10.002. Epub 2005 Oct 26.
Promoter of the Prnd gene coding for the Prion-like protein Doppel contains two critical cis-regulatory elements, NF-Y consensus motif and canonical E-box. Here, we studied a role of the upstream stimulatory factor (USF) in the E-box-mediated activation of Prnd transcription. Co-expression of USF-1 with the luciferase reporter gene driven by the -185/+27 Prnd promoter fragment resulted in several fold increase of the luciferase activity. Conversely, mutations within the E-box led to a significantly reduced Prnd promoter activation. USF-1 binding was supported by the gel shift assay, supershift with USF-1 antibody and UV cross-linking. The activation capacity of the related USF-2, c-Myc and HIF-2alpha proteins was lower compared to USF-1 suggesting that USF-1 is the major E-box-binding transcription factor regulating the Prnd promoter.
编码类朊病毒蛋白多普蛋白（Doppel）的Prnd基因启动子包含两个关键的顺式调控元件，即核转录因子Y（NF-Y）共有基序和典型的E盒。在此，我们研究了上游刺激因子（USF）在E盒介导的Prnd转录激活中的作用。USF-1与由-185/+27 Prnd启动子片段驱动的荧光素酶报告基因共表达，导致荧光素酶活性增加了几倍。相反，E盒内的突变导致Prnd启动子激活显著降低。凝胶迁移实验、用USF-1抗体进行的超迁移实验和紫外线交联实验均证实了USF-1的结合。与USF-1相比，相关的USF-2、c-Myc和低氧诱导因子-2α（HIF-2α）蛋白的激活能力较低，这表明USF-1是调节Prnd启动子的主要E盒结合转录因子。
