Dean D C, Gope R, Knoll B J, Riser M E, O'Malley B W
J Biol Chem. 1984 Aug 25;259(16):9967-70.
We have previously transferred an ovalbumin-beta-globin fusion gene (ovalglobin) into primary cultures of chick oviduct cells and demonstrated that an ovalbumin gene 5'-flanking sequence between -221 and -95 is necessary for progesterone-mediated transcriptional induction (Dean, D. C., Knoll, B. J., Riser, M. E., and O'Malley, B. W. (1983) Nature (Lond.) 305, 551-554). Here we compare 5'-flanking sequences required for induction of the ovalglobin gene by 17 beta-estradiol and progesterone. The early gene of simian virus 40 was inserted into the same plasmid as the ovalbumin fusion gene to serve as an internal control. Since transcription of the viral early gene was unaffected by the presence of steroid hormone or deletions in the ovalbumin gene 5'-flanking region, the level of its transcripts could be monitored as a reference standard for ovalglobin transcription. Ovalglobin transcripts initiated principally from the ovalbumin cap site in the presence or absence of progesterone and 17 beta-estradiol. Deletion of 5'-flanking sequences to -197 had little effect on the induction with either hormone, while successive deletions to -180, -161, and -143 resulted in a gradual decrease in the level of induction. Deletion to -95 eliminated the induction. The results of this study indicate that DNA control elements for regulation of the ovalbumin gene by estrogen and progesterone either overlap directly or are clustered in close proximity in the 5'-flanking region near the ovalbumin gene promoter.
我们之前已将一个卵清蛋白-β-珠蛋白融合基因(卵珠蛋白)转入鸡输卵管细胞原代培养物中,并证明在-221至-95之间的卵清蛋白基因5'侧翼序列对于孕酮介导的转录诱导是必需的(迪恩,D.C.,诺尔,B.J.,里泽,M.E.,和奥马利,B.W.(1983年)《自然》(伦敦)305,551 - 554)。在此我们比较17β-雌二醇和孕酮诱导卵珠蛋白基因所需的5'侧翼序列。将猴病毒40的早期基因插入与卵清蛋白融合基因相同的质粒中作为内部对照。由于病毒早期基因的转录不受类固醇激素的存在或卵清蛋白基因5'侧翼区域缺失的影响,其转录本水平可作为卵珠蛋白转录的参考标准进行监测。无论有无孕酮和17β-雌二醇,卵珠蛋白转录本主要从卵清蛋白帽位点起始。将5'侧翼序列缺失至-197对两种激素的诱导作用影响不大,而连续缺失至-180、-161和-143导致诱导水平逐渐降低。缺失至-95消除了诱导作用。本研究结果表明,雌激素和孕酮调控卵清蛋白基因的DNA控制元件要么直接重叠,要么在卵清蛋白基因启动子附近的5'侧翼区域紧密聚集。