Molecular mechanisms of nucleic acid machines

核酸机器的分子机制

• 批准号: 10624842
• 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/10624842
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Base Pairing; Binding; Cell division; Complex; Cryoelectron Microscopy; DNA; DNA biosynthesis; Data; Disease; Enzymes; Eukaryotic Cell; Event; Genetic Materials; HIV; Hepatitis C virus; Human; In Vitro; Knowledge; 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复制是所有生物的基本过程，以精确复制遗传物质 细胞分裂。该过程的核心是一种解旋酶，该酶使用ATP-Hydrolsys分离碱基对 DNA允许聚合酶获得合成的互补链，并驱动 沿着DNA的复制机械。在人和其他真核细胞中，解旋酶发动机是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差距 MCM蛋白与ATP化合物和DNA相互作用时的机理图片。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了研究 晶体学和冷冻电子显微镜以及研究其功能的体外方法和 互动。死盒RNA解旋酶DDX3X与RNA的许多方面有关 加工并与几种人类疾病有关，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差距 具有功能状态的RNA和ATP化合物。大量的初步数据已经 为该项目获得的，其中包括鉴定具有RNA-的DDX3X的最小片段 刺激ATPase活性及其与相关底物结合的晶体结构。

项目成果

Two Distinct Modes of DNA Binding by an MCM Helicase Enable DNA Translocation.

• DOI: 10.3390/ijms232314678
• 发表时间: 2022-11-24
• 期刊: International journal of molecular sciences
• 影响因子: 5.6

Structure of a dimer of the Sulfolobus solfataricus MCM N-terminal domain reveals a potential role in MCM ring opening.

硫磺硫化叶菌 MCM N 末端结构域二聚体的结构揭示了 MCM 开环中的潜在作用。

• DOI: 10.1107/s2053230x21005331
• 发表时间: 2021
• 期刊: Acta crystallographica. Section F, Structural biology communications
• 作者: Meagher,Martin;Spence,MadisonN;Enemark,EricJ
• 通讯作者: Enemark,EricJ