Structural and biophysical studies of proteins, nucleic acids, and their complexes

蛋白质、核酸及其复合物的结构和生物物理研究

• 批准号: 10623936
• 负责人: Alfonso Mondragon
• 金额: $ 58.29万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10623936
• 关键词: Affect; Aminoacylation; Antibiotics; Architecture; Binding; Biological; Biological Process; Cell physiology; Complex; DNA; Elements; Evolution; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Life; Ligands; Malignant Neoplasms; Messenger RNA; Nucleic Acids; Property; Proteins; RNA; Regulator Genes; Research; Research Design; Role; Signal Transduction; Structure; Therapeutic Agents; Topoisomerase; Transfer RNA; Translations; Untranslated RNA; Untranslated Regions; Work; biophysical analysis; chemotherapeutic agent; design; macromolecule; novel therapeutics; single molecule; small molecule

Project Summary The central theme of this proposal is structural and mechanistic studies of nucleic acids, proteins, and their interactions. The two broad topics that will serve as foci for our future research are: 1) the structure and mechanism of topoisomerases, and 2) the structure and mechanism of non-coding RNA molecules, particularly T-box riboswitches. The aim of our work is to discover and explore paradigms that help to illuminate the structure and mechanism of biological molecules. Topoisomerases are ubiquitous proteins found across all three domains of life. They are involved in several cellular processes and the importance of their cellular role is underscored by the fact that they are the target of several cancer chemotherapeutic agents and antibiotics. The study of the structure, mechanism, and function of topoisomerases not only furthers our understanding at many different levels of proteins that interact with DNA and alter its topological properties, but also provides important information to aid in the design of new therapeutic agents. We will continue our comprehensive studies of topoisomerases with an emphasis on three large and complementary sub-projects: i) Single molecule studies of type IA topoisomerases, ii) Structural studies of topoisomerases on natural substrates, and iii) Structural studies of gyrase in complex with mini- circle DNA. These sub-projects will help answer crucial questions that are needed to advance the field to the next level. Riboswitches are ligand-specific cis-regulatory RNA elements found in untranslated regions of mRNAs that respond to defined external signals to affect transcription or translation of downstream genes. Bacterial T- box riboswitches represent a unique class of riboswitches that do not bind small molecule ligands, instead they recognize and bind tRNA molecules and sense directly their aminoacylation state. They represent an excellent paradigm to understand RNA structure and architecture, recognition of RNA molecules by other RNA molecules, RNA-based gene regulation, and the evolution of gene regulatory function from the ancient RNA world. We will continue and expand our studies of T-box riboswitches by concentrating in three subprojects: i) Single molecule studies of translational T-box riboswitches, ii) Structural and single molecule studies of typical T-box riboswitches, and iii) Structural studies of tandem T-box riboswitches. These three sub-projects will advance our understanding of T-box riboswitches and non-coding RNA molecules, molecules that are still not fully understood at the atomic level.

项目概要 该提案的中心主题是核酸、蛋白质和生物体的结构和机制研究 他们的互动。将作为我们未来研究重点的两个广泛主题是：1）结构和 拓扑异构酶的机制，以及2）非编码RNA分子的结构和机制，特别是 T 盒核糖开关。我们工作的目的是发现和探索有助于阐明 生物分子的结构和机制。 拓扑异构酶是在生命的所有三个领域中普遍存在的蛋白质。他们参与 一些细胞过程及其细胞作用的重要性通过以下事实得到强调： 几种癌症化疗药物和抗生素的靶点。结构、机制和作用的研究 拓扑异构酶的功能不仅进一步加深了我们对相互作用的许多不同水平的蛋白质的理解 与 DNA 结合并改变其拓扑特性，而且还提供重要信息来帮助设计 新的治疗剂。我们将继续对拓扑异构酶进行全面研究，重点是 三个大型且互补的子项目：i) IA 型拓扑异构酶的单分子研究，ii) 结构 天然底物上拓扑异构酶的研究，以及 iii) 旋转酶与微型复合物的结构研究 环状DNA。这些子项目将有助于回答将该领域推进到下一个阶段所需的关键问题。 等级。 核糖开关是在 mRNA 非翻译区域中发现的配体特异性顺式调节 RNA 元件 响应定义的外部信号以影响下游基因的转录或翻译。细菌T- 盒式核糖开关代表了一类独特的核糖开关，它们不结合小分子配体，而是 识别并结合 tRNA 分子并直接感知其氨酰化状态。他们代表着优秀的 理解 RNA 结构和结构、RNA 分子被其他 RNA 分子识别的范式， 基于RNA的基因调控，以及来自古代RNA世界的基因调控功能的进化。我们将 通过集中于三个子项目继续并扩展我们对 T 盒核糖开关的研究： i) 单分子 翻译T-盒核糖开关的研究，ii) 典型T-盒的结构和单分子研究 核糖开关，以及 iii) 串联 T 盒核糖开关的结构研究。这三个子项目将推进 我们对T-box核糖开关和非编码RNA分子的理解，这些分子仍然不完全 在原子水平上理解。