Pugh C W, Tan C C, Jones R W, Ratcliffe P J
Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford, United Kingdom.
Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10553-7. doi: 10.1073/pnas.88.23.10553.
Erythropoietin, the major hormone controlling red-cell production, is regulated in part through oxygen-dependent changes in the rate of transcription of its gene. Using transient transfection in HepG2 cells, we have defined a DNA sequence, located 120 base pairs 3' to the poly(A)-addition site of the mouse erythropoietin gene, that confers oxygen-regulated expression on a variety of heterologous promoters. The sequence has the typical features of a eukaryotic enhancer. Approximately 70 base pairs are necessary for full activity, but reiteration restores activity to shorter inactive sequences. This enhancer operates in HepG2 and Hep3B cells, but not in Chinese hamster ovary cells or mouse erythroleukemia cells, and responds to cobalt but not to cyanide or 2-deoxyglucose, thus reflecting the physiological control of erythropoietin production accurately.
促红细胞生成素是控制红细胞生成的主要激素，其部分通过其基因转录速率的氧依赖性变化来调节。利用HepG2细胞中的瞬时转染，我们确定了一个DNA序列，该序列位于小鼠促红细胞生成素基因的多聚腺苷酸加尾位点下游120个碱基对处，它能赋予多种异源启动子氧调节表达。该序列具有真核增强子的典型特征。大约70个碱基对是充分活性所必需的，但重复能使较短的无活性序列恢复活性。这种增强子在HepG2和Hep3B细胞中起作用，但在仓鼠卵巢细胞或小鼠红白血病细胞中不起作用，并且对钴有反应，但对氰化物或2-脱氧葡萄糖没有反应，从而准确地反映了促红细胞生成素产生的生理控制。
