Firth J D, Ebert B L, Pugh C W, Ratcliffe P J
Erythropoietin Group, Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford, England.
Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6496-500. doi: 10.1073/pnas.91.14.6496.
Production of the glycoprotein hormone erythropoietin (Epo) in response to hypoxic stimuli is almost entirely restricted to particular cells within liver and kidney, yet the transcriptional enhancer lying 3' to the Epo gene shows activity inducible by hypoxia after transfection into a wide variety of cultured cells. The implication of this finding is that many cells which do not produce Epo contain a similar, if not identical, oxygen-regulated control system, suggesting that the same system is involved in the regulation of other genes. We report that the human phosphoglycerate kinase 1 and mouse lactate dehydrogenase A genes are induced by hypoxia with characteristics which resemble induction of the Epo gene. In each case expression is induced by cobalt, but not by cyanide, and hypoxic induction is blocked by the protein-synthesis inhibitor cycloheximide. We show that the relevant cis-acting control sequences are located in the 5' flanking regions of the two genes, and we define an 18-bp element in the 5' flanking sequence of the phosphoglycerate kinase 1 gene which is both necessary and sufficient for the hypoxic response, and which has sequence and protein-binding similarities to the hypoxia-inducible factor 1 binding site within the Epo 3' enhancer.
对缺氧刺激作出反应时,糖蛋白激素促红细胞生成素(Epo)的产生几乎完全局限于肝脏和肾脏中的特定细胞,然而,位于Epo基因3'端的转录增强子在转染到多种培养细胞后显示出可被缺氧诱导的活性。这一发现意味着许多不产生Epo的细胞含有类似(即便不是相同)的氧调节控制系统,表明同一系统参与了其他基因的调控。我们报告,人磷酸甘油酸激酶1基因和小鼠乳酸脱氢酶A基因可被缺氧诱导,其特征类似于Epo基因的诱导。在每种情况下,表达都是由钴诱导的,而不是由氰化物诱导的,缺氧诱导被蛋白质合成抑制剂环己酰亚胺阻断。我们表明,相关的顺式作用控制序列位于这两个基因的5'侧翼区域,并且我们在磷酸甘油酸激酶1基因的5'侧翼序列中定义了一个18bp的元件,该元件对于缺氧反应既是必需的也是充分的,并且在序列和蛋白质结合方面与Epo 3'增强子内的缺氧诱导因子1结合位点相似。