STRUCTURAL ANALYSIS OF BIOLOGICAL MACROMOLECULES

基因控制的分子基础研究将继续进行 主要关注转录调控的机制和 特定蛋白质/DNA相互作用的化学。我们将继续使用 用高分辨X射线结晶学确定化合物的结构(S) 各个大分子配对及其特定的络合物。这个 从这些结构中得出的推论将受到生物化学的挑战 和基因实验。最后，一种定量的物理化学 对时间平均和动态交互的描述将是 旨在解释以调节蛋白为目标的生物化学 它们的同源DNA元素。 待研究的原核系统有：(1)色氨酸 抑制子/操纵子系统，其结构和化学现在已被了解 更详细地说，-(2)Arg抑制子/操纵子/分解酶系统。 将研究与rna polII调控有关的真核系统。 包括：(1)类固醇受体/反应元件，(2)B-ZIP蛋白 /靶DNA，(3)牛乳头瘤病毒E2/增强子，(4)血管紧张素转换酶-L/UASC，a 金属反应转录因子及其DNA靶点，(5) TFIID/TATA-box，核心聚合酶复合体的关键成员，修复 转录的位置和极性。晶体存在于所有的 除TFIID/TATA-BOX复合体外，其余各系统均有表达。我们的更远距离聚焦 是研究PolII监管组装的相互作用要素。 这将需要紧凑和定义良好的多聚体的工程设计 适合于结晶学的蛋白质/DNA复合体。 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控。在某种程度上， 转录调控的失败或失衡在这种情况下起到了一定作用 病毒感染、恶变和内分泌异常 这里提出的疾病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.