Affinity of interactions between human glucocorticoid receptors and DNA: at physiologic ionic strength, stable binding occurs only with DNAs containing partially symmetric glucocorticoid response elements.
Affinity of interactions between human glucocorticoid receptors and DNA: at physiologic ionic strength, stable binding occurs only with DNAs containing partially symmetric glucocorticoid response elements.
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人糖皮质激素受体与 DNA 之间相互作用的亲和力：在生理离子强度下，仅与含有部分对称糖皮质激素反应元件的 DNA 发生稳定结合。
DOI：
10.1021/bi00480a016
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发表时间：
1990
期刊：
影响因子：
2.9
通讯作者：
Cidlowski,JA
中科院分区：
文献类型：
--
作者：
Tully,DB;Cidlowski,JA
1990年4月13日收到的修订版Mandarin pt摘要：蔗糖密度梯度变化测定法适用于在DNA过量条件下测定人糖皮质激素受体（GR）与DNA之间相互作用的亲和力。进行饱和度分析，以确定活化的人GR与五个DNA片段中的每一个的相互作用的解离常数。在几乎等渗盐条件下，在蔗糖梯度上的GR-DNA复合物的双链反应显示出相似的亲和力，其中GR与含有糖皮质激素反应元件（GRES）的DNA片段的相互作用的解离常数在2-16 nM的范围内，表现出部分二分体对称性。相比之下，GR表现出几乎没有亲和力的非GRE含有DNA或DNA只含有GRE半位点。此外，GR显示出与含有两个部分对称GRE的DNA片段的多位点相互作用的证据，但仅在含有单个部分对称GRE的MMTV LTR DNA片段的一个位点与三个半GRE的簇沿着相互作用。糖皮质激素受体仅对含有特异性部分对称GRES的DNA具有高选择性和亲和力，并进一步表明对这些DNA位点的这种高亲和力可能是足以解释在糖皮质激素应答细胞中观察到的基因表达的选择性调节。
Revised Manuscript Received April 13, 1990 abstract: Sucrose density gradient shift assays were adapted to permit determination of the affinity of interaction between human glucocorticoid receptors (GR) and DNA under conditions of DNA excess. Saturation analyses were performed to ascertain dissociation constants for the interaction of activated human GR with each of five DNA fragments. Centrifugation of GR-DNA complexes on sucrose gradients under nearly isotonic salt conditions revealed similar affinities with dissociation constants in the range of 2-16 nM for GR interaction with DNA fragments containing glucocorticoid response elements (GREs) exhibiting partial dyad symmetry. By contrast, GR exhibited virtually no affinity for non-GRE-containing DNA or for DNA containing only GRE half-sites. Additionally, GR showed evidence of multiple-siteinteraction with a DNA fragment containing two partially symmetric GREs, but interacted at only one site of an MMTV LTR DNA fragment containing a single partially symmetric GRE along with a cluster of three half-GREs.Together these data indicate that under physiologically relevant conditions, glucocorticoid receptors have high selectivity and affinity only for DNA containing specific partially symmetric GREs and further suggest that this high affinity for such DNA sites may be sufficient to account for the selective regulation of gene expression observed in glucocorticoid-responsive cells.