Ohashi Y, Sawada Y, Moriuchi T, Fujinaga K
Department of Molecular Biology, Cancer Research Institute, Sapporo Medical College, Japan.
Biochim Biophys Acta. 1992 Mar 24;1130(2):175-81. doi: 10.1016/0167-4781(92)90525-5.
The proliferating cell nuclear antigen (PCNA), highly conserved among eukaryotes, is an auxiliary factor for DNA polymerase delta. In this report we sequenced 1560 nucleotides (nt) of the 5' flanking region of the rat PCNA gene and located the transcription initiation site. The sequence contains 1435 nt upstream of the cap site and promotes transcription of a linked heterologous reporter gene in rat, mouse and human cells. Transient expression assays using a series of 5' deletion mutants revealed that 240 nt of the upstream sequence are sufficient for full promoter activity. Three GC boxes and several other binding sites of transcription factors were observed, but neither a TATA nor a CCAAT sequence was found in this region. The results also suggested the existence of a negative regulatory element(s) between -968 and -691. Cotransfection with early region 1 (E1) genes of human adenoviruses activated the expression of the reporter gene, suggesting that an E1-responsive element is located at the proximal promoter region within 81 nt upstream of the transcription initiation site.
增殖细胞核抗原（PCNA）在真核生物中高度保守，是DNA聚合酶δ的辅助因子。在本报告中，我们对大鼠PCNA基因5'侧翼区的1560个核苷酸（nt）进行了测序，并确定了转录起始位点。该序列在帽位点上游包含1435 nt，并能促进大鼠、小鼠和人类细胞中连接的异源报告基因的转录。使用一系列5'缺失突变体进行的瞬时表达分析表明，上游序列的240 nt足以实现完整的启动子活性。观察到三个GC盒和几个其他转录因子结合位点，但在该区域未发现TATA序列或CCAAT序列。结果还表明在-968至-691之间存在一个负调控元件。与人腺病毒早期区域1（E1）基因共转染可激活报告基因的表达，表明E1反应元件位于转录起始位点上游81 nt内的近端启动子区域。
