Berry M, Nunez A M, Chambon P
Unité 184 de Biologie Moléculaire et de Génie Génétique de l'Institut National de la Santé et de la Recherche Médicale, Faculté de Médecine, Strasbourg, France.
Proc Natl Acad Sci U S A. 1989 Feb;86(4):1218-22. doi: 10.1073/pnas.86.4.1218.
Using chimeric recombinants transfected into HeLa cells and a transient expression assay, we demonstrate that the 5'-flanking region of the pS2 gene from position -428 to position -324 exhibits both constitutive and estrogen-inducible enhancer activity. The estrogen-inducible activity, but not the constitutive activity, was inhibited by antiestrogens. ICI 164,384 behaved as a pure antagonist, whereas hydroxytamoxifen was a partial agonist-antagonist. The estrogen-responsive element of the pS2 gene has been narrowed down by site-directed deletion mutagenesis to a 13-base-pair (position -405 to position -393) imperfectly palindromic sequence, which in isolation can confer estrogen inducibility to the heterologous rabbit beta-globin gene promoter. On the other hand, the sequences responsible for the constitutive enhancer activity are spread over the entire region.
通过将嵌合重组体转染到HeLa细胞中并进行瞬时表达分析,我们证明了pS2基因从-428位到-324位的5'侧翼区域表现出组成型和雌激素诱导型增强子活性。抗雌激素抑制了雌激素诱导活性,但不抑制组成型活性。ICI 164,384表现为纯拮抗剂,而羟基他莫昔芬是部分激动剂-拮抗剂。通过定点缺失诱变,pS2基因的雌激素反应元件已缩小到一个13个碱基对(-405位到-393位)的不完全回文序列,该序列单独即可赋予异源兔β-珠蛋白基因启动子雌激素诱导性。另一方面,负责组成型增强子活性的序列分布在整个区域。