RECEPTOR BINDING FACTOR FOR THE AH-RECEPTOR

AH 受体的受体结合因子

• 批准号: 2377904
• 负责人: MARY F RUH
• 金额: $ 18.46万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2377904
• 关键词: DNA binding protein; SDS polyacrylamide gel electrophoresis; affinity chromatography; aminoacid analyzer; animal tissue; aromatic hydrocarbon receptor; chromatin; dioxins; genetic transcription; laboratory rabbit; liver; nuclear matrix; nucleic acid sequence; polymerase chain reaction; protein reconstitution; receptor binding; receptor sensitivity; southern blotting; steroid hormone receptor; uterus; western blottings

In order to better understand the cellular mechanisms involved in the diverse effects of the environmental contaminant, dioxin, we will study specific receptor binding factors (RBFs) for the Ah receptor in chromatin. The differential sensitivity of various animal species and tissues to dioxin may be explained, in part, by the absence or presence of certain AhRBFs in the nuclear matrix. In addition, the antiestrogenic actions of dioxin may be explained by interaction of the Ah receptor with certain uterine estrogen RBFs. These studies will also address the poor correlation between biological effectiveness of some compounds and their in vitro binding affinity for the Ah receptor or for receptor complex binding to dioxin response elements. We must first purify select liver and uterine AhRBFs. Determination of their amino acid sequences will allow for comparisons with other DNA binding proteins, especially RBFs for steroid hormones. Preparation of antibodies to these AhRBFs will allow us to determine if these chromosomal proteins are associated with the nuclear matrix in a species and tissue specific manner. Studies are also proposed to investigate the role of AhRBFs in gene transcription utilizing both transient and stable transfection experiments. We will determine if and where AhRBFs are associated with known dioxin-inducible genes, such as CYP1A1 or CYP1A2 and also determine the consensus DNA sequence to which these AhRBFs bind.

为了更好地理解参与的细胞机制 环境污染物二恶英的多种影响，我们将研究 染色质中 Ah 受体的特异性受体结合因子 (RBF)。 不同动物种类和组织的敏感性不同 二恶英的部分原因可能是由于某些物质的存在或不存在 核基质中的 AhRBF。 此外，抗雌激素作用 二恶英可能是通过 Ah 受体与某些特定物质的相互作用来解释的。 子宫雌激素 RBF。 这些研究还将针对贫困人口 某些化合物的生物有效性与其作用之间的相关性 Ah 受体或受体复合物的体外结合亲和力 与二恶英反应元件结合。 首先要净化选择肝脏 和子宫 AhRBF。 确定它们的氨基酸序列将 允许与其他 DNA 结合蛋白进行比较，尤其是 RBF 类固醇激素。 制备这些 AhRBF 的抗体将使我们能够 以确定这些染色体蛋白是否与核相关 基质以物种和组织特定的方式。 还提出了研究 利用两者研究 AhRBF 在基因转录中的作用 瞬时和稳定转染实验。 我们将确定是否以及 其中 AhRBF 与已知的二恶英诱导基因相关，例如 CYP1A1 或 CYP1A2 并确定共有 DNA 序列 这些 AhRBF 结合。