STRUCTURAL ANALYSIS OF BIOLOGICAL MACROMOLECULES

生物大分子结构分析

• 批准号: 3484183
• 负责人: PAUL B SIGLER
• 金额: $ 32.42万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-01-01 至 1996-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3484183
• 关键词: DNA binding protein; X ray crystallography; bacterial genetics; bovine papillomavirus; chemical structure function; conformation; gel electrophoresis; gene expression; genetic enhancer element; genetic operator element; genetic regulatory element; genetic transcription; molecular site; nucleic acid sequence; nucleic acid structure; oncogenes; protein structure; site directed mutagenesis; stereochemistry; steroid hormone receptor; stoichiometry; thermodynamics; transcription factor

Research will proceed on the molecular basis of genetic control with primary focus on the mechanisms of transcriptional regulation and the chemistry of specific protein/DNA interactions. We will continue to use high resolution X-ray crystallography to establish the structure(s) of the individual macromolecular partners and their specific complexes. The inferences drawn from these structures will be challenged by biochemical and genetic experiments. Finally, a quantitative physical chemical description of the interaction both time averaged and dynamic, will be developed to explain the biochemistry that targets regulatory proteins to their cognate DNA elements. The prokaryotic systems to be studied are: (1) the trp repressor/operator system whose structure and chemistry is now understood in great detail and, - (2) the arg repressor/operator/resolvase system. Eukaryotic systems involved in RNA polII regulation will be studied including: (1) the steroid receptor/response element, (2) B-zip proteins /target DNA, (3) E2/enhancer of bovine papillomavirus, (4) ACE- l/UASc, a metalresponsive transcription factor and its DNA target, (5) TFIID/TATA-box, a key member of the core polymerase complex that fixes the position and polarity of transcription. Cocrystals exist for all of the above systems except TFIID/TATA-box complex. Our longer range focus is to study the interacting elements of the PolII regulatory assembly. This will require the engineering of compact and well defined multimeric protein/DNA complexes suitable for crystallography. The differentiation, growth and metabolism of healthy cells depends on the precise regulation of gene expression. To the extent that failures or imbalances of transcriptonal control play a role in such abnormalities as viral infection, malignant transformation, and endocrine disorders the studies proposed here are of fundamental biomedical importance.

研究将以遗传控制的分子基础进行 主要关注转录调节机制和 特定蛋白/DNA相互作用的化学。 我们将继续使用 高分辨率X射线晶体学以建立结构 单个大分子伴侣及其特定的络合物。 这 从这些结构中得出的推论将受到生化的挑战 和基因实验。 最后，一种定量的物理化学 相互作用的描述平均和动态的时间将是 开发以解释针对调节蛋白的生物化学 他们的同源DNA元素。 要研究的核系统是：（1）TRP 现在了解其结构和化学的阻遏物/操作员系统 详细详细介绍 - （2）ARG阻遏物/操作员/分辨率系统。 将研究参与RNA Polii调节的真核系统 包括：（1）类固醇受体/反应元件，（2）B-ZIP蛋白 /靶DNA，（3）E2/牛乳头瘤病毒的增强子，（4）Ace-L/UASC，A 金属响应转录因子及其DNA靶标，（5） tfiid/tata-box，修复的核心聚合酶复合酶的关键成员 转录的位置和极性。 所有共同性存在 除TFIID/TATA-BOX复合物外，上述系统。 我们较长的范围重点 是研究polii调节组装的相互作用元素。 这将需要紧凑且定义良好的多聚体的工程 蛋白质/DNA复合物适用于晶体学。 健康细胞的分化，生长和代谢取决于 关于基因表达的精确调节。 在某种程度上 转录控制的失败或失衡在这种情况下起作用 异常作为病毒感染，恶性转化和内分泌 疾病此处提出的研究是基本的生物医学 重要性。