Scheidereit C, Geisse S, Westphal H M, Beato M
Nature. 1983;304(5928):749-52. doi: 10.1038/304749a0.
Glucocorticoids are known to induce the transcription of integrated proviral mouse mammary tumour virus (MMTV) genes in a variety of cell lines derived from mouse mammary tumours. Chimaeric genes in which selectable markers are linked to the long terminal repeat (LTR) region of MMTV can be induced by the synthetic glucocorticoid dexamethasone after introduction into mouse fibroblasts. This suggests that the regulatory elements required for hormonal induction are located within the cloned LTR fragments. The idea is supported by the observation that glucocorticoid receptors bind to certain cloned fragments of MMTV DNA in vitro. Using filter binding studies and monoclonal antibodies to the glucocorticoid receptor we have now delimited the receptor binding region to a DNA segment of 152 base pairs (bp) that has been shown to be relevant for hormonal induction. In nuclease protection experiments we have identified partially homologous receptor binding sequences located in this region, all of which share the hexanucleotide 5'-TGTTCT-3'.
已知糖皮质激素可在源自小鼠乳腺肿瘤的多种细胞系中诱导整合的前病毒小鼠乳腺肿瘤病毒(MMTV)基因的转录。将可选择标记与MMTV的长末端重复序列(LTR)区域相连的嵌合基因在导入小鼠成纤维细胞后,可被合成糖皮质激素地塞米松诱导。这表明激素诱导所需的调控元件位于克隆的LTR片段内。这一观点得到了以下观察结果的支持:糖皮质激素受体在体外可与MMTV DNA的某些克隆片段结合。利用滤膜结合研究以及针对糖皮质激素受体的单克隆抗体,我们现已将受体结合区域界定为一个152个碱基对(bp)的DNA片段,该片段已被证明与激素诱导相关。在核酸酶保护实验中,我们在该区域鉴定出了部分同源的受体结合序列,所有这些序列都共享六核苷酸5'-TGTTCT-3'。