Sun L H, Pfendner E G, Senior M B, Frankel F R
Mol Cell Endocrinol. 1983 Jun;30(3):267-78. doi: 10.1016/0303-7207(83)90063-1.
MCF-7 cells contain progesterone, estradiol and glucocorticoid receptors. Following addition of these hormones to the growth medium of the cells, hormone-receptor complexes were found to sediment with chromatin fragments produced by trace digestion with micrococcal nuclease. The binding in all cases could be competed by excess unlabeled hormone. In each case the fragments with which the hormone-receptor complexes were associated tended to be smaller than the bulk chromatin fragments, indicating a greater sensitivity of those chromatin regions to the nuclease. The mononucleosomes released by more extensive digestion with micrococcal nuclease contained different amounts of each of the three hormone-receptor complexes. Progesterone could usually be detected on mononucleosomes only after very brief sedimentation analyses, whereas glucocorticoid- and estradiol-labeled mononucleosomes were stable during long centrifugations. Comparison of glucocorticoid- and estradiol-labeled mononucleosomes indicated that their sedimentation rates differed from one another and from bulk nucleosomes. Estradiol nucleosomes from MCF-7 cells and rat uterus (Senior and Frankel, 1978) sediment significantly faster than bulk nucleosomes, while glucocorticoid nucleosomes from MCF-7 cells and rat hepatoma cells sediment with, or even fractionally slower than, bulk nucleosomes.
MCF - 7细胞含有孕酮、雌二醇和糖皮质激素受体。在向细胞生长培养基中添加这些激素后,发现激素 - 受体复合物与微球菌核酸酶微量消化产生的染色质片段一起沉降。在所有情况下,过量未标记的激素都能竞争这种结合。在每种情况下,与激素 - 受体复合物相关的片段往往比大部分染色质片段小,这表明那些染色质区域对核酸酶更敏感。用微球菌核酸酶进行更广泛消化释放的单核小体含有不同量的三种激素 - 受体复合物。只有在非常短暂的沉降分析后,通常才能在单核小体上检测到孕酮,而糖皮质激素和雌二醇标记的单核小体在长时间离心过程中是稳定的。糖皮质激素和雌二醇标记的单核小体的比较表明,它们的沉降速率彼此不同,也与大部分核小体不同。来自MCF - 7细胞和大鼠子宫的雌二醇核小体(Senior和Frankel,1978)沉降速度明显快于大部分核小体,而来自MCF - 7细胞和大鼠肝癌细胞的糖皮质激素核小体与大部分核小体一起沉降,甚至沉降速度略慢于大部分核小体。