Choi T, Cho N, Oh Y, Yoo M, Matsukage A, Ryu Y, Han K, Yoon J, Baek K
Department of Molecular Biology, College of Natural Science, Pusan National University, Pusan 609-735, South Korea.
FEBS Lett. 2000 Oct 13;483(1):71-7. doi: 10.1016/s0014-5793(00)02085-8.
The TATA box binding protein (TBP) is a general transcription factor required for initiation by all three eukaryotic RNA polymerases. Previously, we found that the promoter region of the Drosophila melanogaster TBP gene contains three sequences similar to the DNA replication-related element (DRE) (5'-TATCGATA). In the present study, we found that the DRE-like sequences are also present in the promoter of the Drosophila virilis TBP gene, suggesting a role for these sequences in TBP expression. Band mobility shift assays revealed that oligonucleotides containing sequences similar to the DRE of D. melanogaster TBP gene promoter form specific complexes with a factor in a Kc cell nuclear extract and with recombinant DRE-binding factor (DREF). Furthermore, these complexes were either supershifted or diminished by monoclonal antibodies to DREF. Transient luciferase assays demonstrated that induction of mutations in two DRE-related sequences at positions -223 and -63 resulted in an extensive reduction of promoter activity. Thus, the DRE-DREF system appears to be involved in the expression of the D. melanogaster TBP gene.
TATA 盒结合蛋白（TBP）是所有三种真核生物 RNA 聚合酶起始转录所必需的通用转录因子。此前，我们发现黑腹果蝇 TBP 基因的启动子区域包含三个与 DNA 复制相关元件（DRE）（5'-TATCGATA）相似的序列。在本研究中，我们发现类似 DRE 的序列也存在于粗壮果蝇 TBP 基因的启动子中，这表明这些序列在 TBP 表达中发挥作用。凝胶迁移率变动分析显示，含有与黑腹果蝇 TBP 基因启动子 DRE 相似序列的寡核苷酸与 Kc 细胞核提取物中的一种因子以及重组 DRE 结合因子（DREF）形成特异性复合物。此外，这些复合物被抗 DREF 的单克隆抗体超迁移或减弱。瞬时荧光素酶分析表明，在 -223 和 -63 位置的两个 DRE 相关序列中引入突变会导致启动子活性大幅降低。因此，DRE-DREF 系统似乎参与了黑腹果蝇 TBP 基因的表达。
