Renkawitz R, Danesch U, Matthias P, Schütz G
J Steroid Biochem. 1984 Jan;20(1):99-104. doi: 10.1016/0022-4731(84)90195-x.
To study the mechanism of steroid induced transcriptional control we have introduced recombinants of the chicken lysozyme gene and the rat tryptophan oxygenase (TO) gene into heterologous and homologous cells. To monitor the activity of the TO-promoter, 1.9 kb of the TO 5'-flanking sequences were fused with sequences coding for the bacterial enzyme chloramphenicol acetyltransferase (CAT). Upon transfer into mouse L-cells the transient expression of the TO-CAT recombinant was found to be inducible by dexamethasone. Transient expression of chicken lysozyme gene recombinants after introduction into various cell types could only be detected in chicken oviduct cells, or in conjunction with SV-40 enhancer sequences in human cells. The recombinant gene used in oviduct cells was a fusion between the lysozyme promoter, including 1.4 kb of upstream sequences, and the coding region of the gene for SV 40 T-antigen (plys-T). The expression in oviduct cells was stimulated by dexamethasone or progesterone, whereas SV 40 enhanced expression of lysozyme sequences in human cells could not be regulated by steroids. Using several deletion mutants, a region between -220 bp and -140 bp upstream of the cap site was found to be essential for both regulation by glucocorticoids as well as by progesterone.
为了研究类固醇诱导的转录调控机制，我们将鸡溶菌酶基因和大鼠色氨酸加氧酶（TO）基因的重组体导入了异源和同源细胞中。为了监测TO启动子的活性，将1.9 kb的TO 5'侧翼序列与编码细菌氯霉素乙酰转移酶（CAT）的序列融合。转入小鼠L细胞后，发现TO-CAT重组体的瞬时表达可被地塞米松诱导。将鸡溶菌酶基因重组体导入各种细胞类型后，仅在鸡输卵管细胞中或与人细胞中的SV-40增强子序列结合时才能检测到瞬时表达。在输卵管细胞中使用的重组基因是溶菌酶启动子（包括1.4 kb上游序列）与SV 40 T抗原（plys-T）基因编码区的融合体。输卵管细胞中的表达受地塞米松或孕酮刺激，而人细胞中SV 40增强的溶菌酶序列表达不受类固醇调节。使用多个缺失突变体，发现帽位点上游-220 bp至-140 bp之间的区域对于糖皮质激素和孕酮的调节均至关重要。
