Pavlik E J, Katzenellenbogen B S
Mol Cell Endocrinol. 1982 Apr;26(1-2):201-16. doi: 10.1016/0303-7207(82)90017-x.
The intranuclear locations at which rat uterine estrogen receptors interact with chromatin have been probed using digestions performed with DNAase I and micrococcal nuclease. Exposure to nuclease has been controlled to effect limited to extensive digestion of nuclear DNA under conditions which maintain the integrity of the [3H] estradiol-receptor complex. The effect of divalent cation concentration on the release of estrogen receptors fron nuclease-treated chromatin was examined and found to be of consequence above 2 mM. Exposure to nuclease released nuclear estrogen receptors from chromatin, with DNAase I being more efficient than micrococcal nuclease in mediating this release. The release of the bulk of nuclear estrogen receptors closely paralleled the nuclease-mediated digestion of chromatin DNA. At 1 h after exposure to estrogen, substantial quantities of uterine estrogen receptors (80-90%) were distributed in chromatin fractions which, on the basis of fractionation terminology, have been termed 'transcriptionally inactive' by convention. Enrichment of estrogen receptors in chromatin which has been termed ' transcriptionally active' only occurred with 10-20% of the estrogen receptors. Hence, our findings support a model where, at early times after estrogen exposure, receptors from the rat uterus are enriched to only a minor extent in chromatin to which 'transcriptional activity' is generally assigned while the bulk of receptors are localized in chromatin which is generally considered 'transcriptionally inactive'.
利用脱氧核糖核酸酶I(DNAase I)和微球菌核酸酶进行消化,已对大鼠子宫雌激素受体与染色质相互作用的核内位置进行了探究。在维持[3H]雌二醇-受体复合物完整性的条件下,已对核酸酶的暴露进行控制,以实现对核DNA从有限消化到广泛消化的效果。研究了二价阳离子浓度对核酸酶处理的染色质中雌激素受体释放的影响,发现当浓度高于2 mM时具有重要影响。核酸酶处理可使染色质释放核雌激素受体,在介导这种释放方面,DNAase I比微球菌核酸酶更有效。大部分核雌激素受体的释放与核酸酶介导的染色质DNA消化密切平行。在暴露于雌激素1小时后,大量子宫雌激素受体(80 - 90%)分布在染色质组分中,根据分级分离术语,传统上这些组分被称为“转录无活性”。仅10 - 20%的雌激素受体出现在被称为“转录活性”的染色质中,从而实现了雌激素受体的富集。因此,我们的研究结果支持这样一种模型:在雌激素暴露后的早期,大鼠子宫中的受体仅在一定程度上富集于通常被认为具有“转录活性”的染色质中,而大部分受体则定位于通常被认为“转录无活性”的染色质中。