Takahashi Y, Yamaguchi M, Hirose F, Kobayashi J, Miyajima S, Matsukage A
Laboratory of Cell Biology, Aichi Cancer Center Research Institute, Nagoya.
J Biochem. 1997 Dec;122(6):1215-23. doi: 10.1093/oxfordjournals.jbchem.a021884.
The promoter region of the Bombyx mori gene encoding the proliferating cell nuclear antigen (PCNA), and its activating factor(s) were analyzed to ascertain similarities with Drosophila regulatory elements. Full promoter activity was established to reside within the region from -466 to +347 base pairs with respect to the transcription initiation site. Within this region, we found a sequence similar to the DNA replication-related element (DRE), 5'-TATCGATA, which is a promoter-activating sequence common to promoters of the Drosophila genes for DNA replication-related factors, including PCNA. A mutation in the DRE-like sequence of the B. mori PCNA gene promoter caused reduction of the promoter activity and also binding to the recombinant Drosophila DRE-binding factor (DREF). Furthermore, a factor(s) binding to the DRE sequence was detected in extracts of B. mori BmN4 cells. Monoclonal antibodies against Drosophila DREF inhibited the binding activity of the factor, as shown by gel mobility shift assays, and allowed specific detection of a 100 kDa protein on immuno Western blot analysis. These results suggest that the B. mori DREF homolog binds to DRE to regulate transcription of the PCNA gene.
对家蚕中编码增殖细胞核抗原（PCNA）的基因的启动子区域及其激活因子进行了分析，以确定其与果蝇调控元件的相似性。已确定完整的启动子活性存在于相对于转录起始位点的-466至+347碱基对区域内。在该区域内，我们发现了一个与DNA复制相关元件（DRE）5'-TATCGATA相似的序列，该序列是果蝇中与DNA复制相关因子（包括PCNA）的基因启动子共有的启动子激活序列。家蚕PCNA基因启动子的DRE样序列中的突变导致启动子活性降低，并且与重组果蝇DRE结合因子（DREF）的结合也减少。此外，在家蚕BmN4细胞提取物中检测到一种与DRE序列结合的因子。凝胶迁移率变动分析表明，针对果蝇DREF的单克隆抗体抑制了该因子的结合活性，并在免疫印迹分析中特异性检测到一种100 kDa的蛋白质。这些结果表明，家蚕DREF同源物与DRE结合以调节PCNA基因的转录。
