NOVEL HOST SYSTEM TO STUDY STEROID HORMONE ACTION

研究类固醇激素作用的新型宿主系统

• 批准号: 3421547
• 负责人: ORLA M. CONNEELY
• 金额: $ 15.61万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-02-12 至 1994-02-11
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3421547
• 关键词: DNA binding protein; Saccharomyces cerevisiae; X ray crystallography; alternatives to animals in research; biological models; biological products; gene expression; genetic manipulation; hormone regulation /control mechanism; mutant; nuclear magnetic resonance spectroscopy; phosphoproteins; progesterone receptors; protein purification; protein structure function; receptor binding; steroid hormone receptor; transcription factor; transposon /insertion element; vitamin D

The cDNA's for several steroid hormone receptors have been cloned and expressed in heterologous cells. The subsequent development of cell transfection assays to analyze receptor mutants has enabled the definition of the functional domains of the receptors. It is clear that they contain distinct domains for DNA and hormone binding. We wish to further define the critical amino acids within these structures required for ligand and DNA binding. This will be accomplished using two complementing strategies. The first is a physical-biochemical (X-ray diffraction and 2D NMR) analysis of the receptor. This requires large quantities of purified protein. For this we propose to use Saccharomyces cerevisiae to overproduce the receptor. The second is to identify mutants of the receptor by designing genetic strategies that will identify key structural components within the molecule. Traditionally endocrinologists have used animal and avian models to study steroid hormone action and as sources of receptor for structural and biochemical analysis. The demand for pure receptor is increasing. Recently however, we have accumulated data to suggest that it may be possible to develop Saccharomyces cerevisiae as a high connectivity model for steroid hormone action. Furthermore, advances in this area we believe, will encourage further use of yeast in related areas, with a commensurate decrease in the use of animal models.

几种类固醇激素受体的cDNA已被克隆，并且 在异源细胞中表达。随后的细胞发展 转染测定分析受体突变体已使该定义 受体的功能域。显然它们包含 DNA和激素结合的不同结构域。我们希望进一步定义 配体和DNA所需的这些结构中的关键氨基酸 结合。这将使用两种补充策略来完成。这 首先是对物理生物化学（X射线衍射和2D NMR）的分析 受体。这需要大量的纯化蛋白质。为了这 我们建议使用酿酒酵母过量生产受体。这 第二是通过设计遗传来鉴定受体的突变体 将确定关键结构组成部分的策略 分子。 传统上，内分泌学家使用动物和鸟类模型来研究 类固醇激素作用，作为结构和结构和 生化分析。对纯受体的需求正在增加。最近 但是，我们积累了数据，以表明可能有可能 开发酿酒酵母作为类固醇的高连通性模型 激素作用。此外，我们相信的这一领域的进步将 鼓励在相关区域进一步使用酵母 减少动物模型的使用。

项目成果

Differential modes of activation define orphan subclasses within the steroid/thyroid receptor superfamily.

不同的激活模式定义了类固醇/甲状腺受体超家族中的孤儿亚类。

• 发表时间: 1992
• 期刊: Gene expression
• 作者: Lydon,JP;Power,RF;Conneely,OM
• 通讯作者: Conneely,OM

A novel, highly regulated, rapidly inducible system for the expression of chicken progesterone receptor, cPRA, in Saccharomyces cerevisiae.

一种新型、高度调控、快速诱导的系统，用于在酿酒酵母中表达鸡孕酮受体 cPRA。

• DOI: 10.1016/0378-1119(92)90706-u
• 发表时间: 1992
• 期刊: Gene
• 影响因子: 3.5
• 作者: Poletti,A;Weigel,NL;McDonnell,DP;Schrader,WT;O'Malley,BW;Conneely,OM
• 通讯作者: Conneely,OM