PDF(Pubmed)
Abstract：
GGAA motifs in the human TP53 and HELB gene promoters play a part in responding to trans‑resveratrol (Rsv) in HeLa S3 cells. This sequence is also present in the 5\'‑upstream region of the human CDC45 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 CDC45 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 CDC45 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.
摘要：
人TP53和HELB基因启动子中的GGAA基序在响应HeLaS3细胞中的反式白藜芦醇（Rsv）中起作用。该序列也存在于人类CDC45基因的5'上游区域，编码CMGDNA解旋酶蛋白复合物的成分。用Rsv（20μM）处理细胞，然后通过定量RT-PCR和蛋白质印迹分析转录本和翻译蛋白，分别。结果表明，处理后CDC45基因和蛋白表达水平均被诱导。为了检查它们是否是由于转录的激活，克隆了CDC45基因的5'上游556bp，并将其插入荧光素酶（Luc）表达载体的多克隆位点。在本研究中,构建了各种缺失/点突变引入的Luc表达质粒，并将其用于瞬时转染测定。结果表明，GGAA基序，包含在推定的RELB蛋白识别序列中，在HeLaS3细胞中响应Rsv的启动子活性中起作用。
