STRUCTURAL ANALYSIS OF BIOLOGICAL MACROMOLECULES

研究将在遗传控制的分子基础上进行， 主要集中在转录调控机制和 特定蛋白质/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，以确定 单个大分子伴侣及其特定复合物。 的 从这些结构中得出的推论将受到生物化学的挑战。 基因实验。 最后，定量物理化学 时间平均的和动态的相互作用的描述，将是 发展来解释靶向调节蛋白的生物化学， 它们的同源DNA元素 待研究的原核系统有：（1）色氨酸 阻遏物/操纵子系统，其结构和化学性质现已被了解 （2）Arg阻遏物/操纵子/解离酶系统。 将研究涉及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/牛乳头瘤病毒增强子，（4）ACE-1/UASc，a 金属应答转录因子及其DNA靶标，（5） TFIID/TATA盒，核心聚合酶复合物的关键成员， 转录的位置和极性。 共晶存在于所有的 上述系统，TFIID/TATA盒复合体除外。 我们的长期目标 是研究PolII调节组件的相互作用元件。 这将需要设计紧凑和明确定义的多聚体 蛋白质/DNA复合物适合晶体学。 健康细胞的分化、生长和代谢取决于 基因表达的精确调控。 的程度 转录控制的失败或不平衡在这种情况下起作用。 病毒感染、恶性转化和内分泌异常 这里提出的研究是基础生物医学的 重要性

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.