Ghazal P, Lubon H, Fleckenstein B, Hennighausen L
Proc Natl Acad Sci U S A. 1987 Jun;84(11):3658-62. doi: 10.1073/pnas.84.11.3658.
The effect of the human cytomegalovirus immediate early region 1 enhancer on transcription was studied in vitro with HeLa cell nuclear extract. Stimulation of in vitro transcription mediated by the enhancer element involves its recognition by specific trans-acting factors present in the nuclear extract. DNase I protection analysis was used to determine at the nucleotide level those enhancer sequences that interact with nuclear factors. At least nine sites of protein-DNA interaction were detected over approximately 400 base pairs of enhancer sequence. The regions of nuclease protection are associated with 21-, 19-, 18-, and 17-base-pair repeat elements as well as with a unique sequence, creating a large nucleoprotein complex. The relationship between the protein binding and the activity of the immediate early region 1 enhancer is discussed.
利用HeLa细胞核提取物在体外研究了人巨细胞病毒立即早期区域1增强子对转录的影响。增强子元件介导的体外转录刺激涉及核提取物中存在的特定反式作用因子对其的识别。DNase I保护分析用于在核苷酸水平确定与核因子相互作用的那些增强子序列。在大约400个碱基对的增强子序列上检测到至少九个蛋白质-DNA相互作用位点。核酸酶保护区域与21、19、18和17个碱基对的重复元件以及一个独特序列相关,形成一个大的核蛋白复合物。讨论了蛋白质结合与立即早期区域1增强子活性之间的关系。