Le Dréan Y, Kern L, Pakdel F, Valotaire Y
Laboratoire de Biologie Moléculaire, URA CNRS 256, Rennes, France.
Mol Cell Endocrinol. 1995 Mar;109(1):27-35. doi: 10.1016/0303-7207(95)03482-m.
The estrogen receptor is a transcription factor that mediates the actions of estrogens which plays a major role in salmonid vitellogenesis. Previously, we cloned and described the estradiol receptor in rainbow trout. To study the transactivation characteristics, the full length cDNA was inserted in an expression vector and tested by transfection on an estrogen-dependent promoter (pERE-TK-CAT). For the first time, direct comparison between the trout receptor (rtER) and human receptor (hER) in an analogous system has been possible. We demonstrate that rtER can, like hER, transactivate transcription in a strictly hormone-dependent manner. The specificity and sensitivity of the rtER response to different steroids have been studied. With rtER, the E2 concentration needed for half maximal activation is 10 times higher than with hER. In addition, we show that rtER has a weak affinity for androgens and transactivation could be induced using high testosterone concentration. Experiments show that both rtER and hER have an equal specificity for estrogens but that the rtER binds its ligand with a lower sensitivity than hER. Several hypotheses concerning the role of main amino acids within the receptor protein are proposed based on the different properties of the receptors and comparison of sequences.
雌激素受体是一种转录因子,介导雌激素的作用,在鲑科鱼类的卵黄生成中起主要作用。此前,我们克隆并描述了虹鳟鱼中的雌二醇受体。为了研究反式激活特性,将全长cDNA插入表达载体,并通过转染雌激素依赖性启动子(pERE-TK-CAT)进行测试。首次在类似系统中对鳟鱼受体(rtER)和人类受体(hER)进行了直接比较。我们证明,rtER与hER一样,能够以严格的激素依赖性方式反式激活转录。研究了rtER对不同类固醇反应的特异性和敏感性。对于rtER,半数最大激活所需的E2浓度比hER高10倍。此外,我们表明rtER对雄激素的亲和力较弱,高浓度睾酮可诱导反式激活。实验表明,rtER和hER对雌激素的特异性相同,但rtER与配体结合的敏感性低于hER。基于受体的不同特性和序列比较,提出了关于受体蛋白中主要氨基酸作用的几种假设。