Tora L, Gaub M P, Mader S, Dierich A, Bellard M, Chambon P
Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Faculté de Médecine, Strasbourg, France.
EMBO J. 1988 Dec 1;7(12):3771-8. doi: 10.1002/j.1460-2075.1988.tb03261.x.
Using a transient co-transfection system we have identified a functional oestrogen responsive element (ERE) in the near vicinity of the TATA box of the chicken ovalbumin gene promoter. Oestrogen induced activation of transcription was observed only in chicken embryo fibroblast cells, and not in human HeLa cells, suggesting the existence of a cell-specific transcription factor(s) that is necessary for the hormonal response. By 5'-deletion mapping and site-directed mutagenesis, a sequence encompassing the more proximal of two GGTCA half-palindromic motifs was identified as a functional ovalbumin gene ERE. Both motifs correspond to oestrogen receptor binding sites in vitro as revealed by DNase I footprinting. Optimal response to the hormonal stimulus apparently requires stereoalignment of the ERE and the TATA box.
利用瞬时共转染系统,我们在鸡卵清蛋白基因启动子的TATA盒附近鉴定出一个功能性雌激素反应元件(ERE)。仅在鸡胚成纤维细胞中观察到雌激素诱导的转录激活,而在人HeLa细胞中未观察到,这表明存在一种细胞特异性转录因子,它是激素反应所必需的。通过5'缺失图谱分析和定点诱变,一个包含两个GGTCA半回文基序中更靠近近端的序列被鉴定为功能性卵清蛋白基因ERE。如DNA酶I足迹分析所示,两个基序在体外均对应雌激素受体结合位点。对激素刺激的最佳反应显然需要ERE和TATA盒的立体排列。