Department of Gene Regulation, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda, Chiba 278‑8510, Japan.
Genomic Medicinal Science, Research Institute for Science and Technology, Tokyo University of Science, Noda, Chiba 278‑8510, Japan.
Mol Med Rep. 2024 Jun;29(6). doi: 10.3892/mmr.2024.13216. Epub 2024 Apr 5.
GGAA motifs in the human and gene promoters play a part in responding to ‑resveratrol (Rsv) in HeLa S3 cells. This sequence is also present in the 5'‑upstream region of the human gene, which encodes a component of CMG DNA helicase protein complex. The cells were treated with Rsv (20 µM), then transcripts and the translated protein were analyzed by quantitative RT‑PCR and western blotting, respectively. The results showed that the gene and protein expression levels were induced after the treatment. To examine whether they were due to the activation of transcription, a 5'‑upstream 556‑bp of the gene was cloned and inserted into a multi‑cloning site of the Luciferase (Luc) expression vector. In the present study, various deletion/point mutation‑introduced Luc expression plasmids were constructed and they were used for the transient transfection assay. The results showed that the GGAA motif, which is included in a putative RELB protein recognizing sequence, plays a part in the promoter activity with response to Rsv in HeLa S3 cells.
人类 和 基因启动子中的 GGAA 基序在 HeLa S3 细胞中对 -resveratrol（Rsv）的反应中起作用。该序列也存在于编码 CMG DNA 解旋酶蛋白复合物组件的人类 基因的 5' 上游区。用 Rsv（20 µM）处理细胞，然后分别通过定量 RT-PCR 和 Western blot 分析转录物和翻译的蛋白质。结果表明，处理后基因和蛋白质表达水平均被诱导。为了检查它们是否是由于转录激活引起的，克隆了 基因的 5' 上游 556bp 并将其插入到 Luciferase（Luc）表达载体的多克隆位点中。在本研究中，构建了各种缺失/点突变引入的 Luc 表达质粒，并用于瞬时转染测定。结果表明，包含在假定的 RELB 蛋白识别序列中的 GGAA 基序在 HeLa S3 细胞中对 Rsv 反应的启动子活性中起作用。
