Molecular mechanisms of nucleic acid machines

核酸机器的分子机制

• 批准号: 10408782
• 负责人: Eric Enemark
• 金额: $ 38万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10408782
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Base Pairing; Cell division; Complex; Cryoelectron Microscopy; Crystallization; DNA; DNA biosynthesis; Data; Disease; Enzymes; Eukaryotic Cell; Event; Genetic Materials; HIV; Hepatitis C virus; Human; In Vitro; Knowledge; Lead; Life; MCM Protein; Malignant Neoplasms; Malignant neoplasm of brain; Methods; Molecular; Nucleic Acids; Organism; Polymerase; Process; Proteins; RNA; RNA Helicase; RNA Processing; Research; Structure; X-Ray Crystallography; cancer therapy; helicase; human disease; medulloblastoma

ABSTRACT DNA replication is a fundamental process for all organisms to precisely duplicate genetic material prior to cell division. Central to the process is a helicase enzyme that uses ATP-hydrolysis to separate base-paired DNA to allow polymerases to gain access to synthesize complementary strands and also to drive the replication machinery along the DNA. In human and other eukaryotic cells, the helicase engine is the 6- protein MCM complex. The proposed research will fill a knowledge gap by providing atomic level mechanism pictures of MCM proteins as they interact with ATP compounds and with DNA. These will be studied at the molecular level by a coordinated approach involving structural studies by X-ray crystallography and cryo-electron microscopy and also in vitro methods to study their functions and interactions. The DEAD-box RNA helicase DDX3X has been implicated in many aspects of RNA processing and is associated with several human diseases, including HIV, HCV, and medulloblastoma. The proposed research will fill a knowledge gap by providing detailed pictures of DDX3X proteins interacting with RNA and ATP compounds in the functional state. A considerable body of preliminary data has been obtained for this project that includes the identification of a minimal fragment of DDX3X that has RNA- stimulated ATPase activity and its crystal structure bound to relevant substrates.

抽象的 DNA复制是所有生物体在形成之前精确复制遗传物质的基本过程。 细胞分裂。该过程的核心是解旋酶，它利用 ATP 水解来分离碱基对 DNA允许聚合酶合成互补链并驱动 沿着DNA的复制机器。在人类和其他真核细胞中，解旋酶引擎是 6- 蛋白质 MCM 复合物。拟议的研究将通过提供原子水平来填补知识空白 MCM 蛋白与 ATP 化合物和 DNA 相互作用的机制图。这些将是 通过涉及 X 射线结构研究的协调方法在分子水平上进行研究 晶体学和冷冻电子显微镜以及体外方法来研究它们的功能和 互动。 DEAD-box RNA 解旋酶 DDX3X 与 RNA 的许多方面有关 处理并与多种人类疾病相关，包括 HIV、HCV 和髓母细胞瘤。这 拟议的研究将通过提供 DDX3X 蛋白质相互作用的详细图片来填补知识空白 RNA 和 ATP 化合物处于功能状态。已获得大量初步数据 该项目获得的信息包括鉴定 DDX3X 的最小片段，该片段具有 RNA- 刺激的 ATP 酶活性及其与相关底物结合的晶体结构。