Deciphering fundamental biological processes involving protein-nucleic acid interactions at the molecular level

破译涉及分子水平上蛋白质-核酸相互作用的基本生物过程

• 批准号: 10319963
• 负责人: Maria Schumacher
• 金额: $ 58.19万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10319963
• 关键词: Actins; Antibiotics; Bacillus subtilis; Biochemical; Biological; Biological Process; Biology; Cryoelectron Microscopy; DNA; DNA Binding; Data; Development; Enzymes; Eukaryota; Genetic; Genetic Transcription; Glutamate-Ammonia Ligase; Goals; Health; Histones; Homeostasis; Homologous Gene; Human; Investigation; Laboratories; Life; Mediating; Microbe; Microscopy; Modality; Modeling; Molecular; Multi-Drug Resistance; Nitrogen; Nucleic Acids; Plasmids; Process; Production; Proteins; Research; Streptomyces; System; Testing; Therapeutic; Tubulin; Walkers; Work; antimicrobial; base; combat; insight; interest; microbial; multidrug tolerance; novel; nucleoside triphosphatase; rational design; segregation

ABSTRACT The central focus of the Schumacher laboratory is to deduce molecular principles that govern fundamental biological processes involving protein-nucleic acid interactions. Our recent work has honed in on transcription networks and DNA segregation in microbes. The latter studies have uncovered key insights into the molecular mechanisms utilized by simplified systems employing actin-like and tubulin-like NTPases to segregate bacterial plasmids. However, the molecular mechanism(s) controlling the most common bacterial segregation systems, the Walker-box based systems, remain unclear and represent a major focus of our research. Strikingly, our recent studies characterizing the first archaeal segregation system revealed that it utilizes a bacterial-like Walker-box NTPase to drive DNA segregation, indicating that Walker-box segregation machineries may be the most ubiquitous type of DNA segregation modules in biology. These investigations also revealed that the archaeal segregation ParB protein harbors a fold similar to CenpA, the histone homolog that mediates DNA segregation in eukaryotes. Thus, these studies uncovered possible evolutionary linkages in segregation machineries between the 3 domains of life. Our most recent work has provided the first molecular views of Walker-box NTPases bound to DNA and ParB. These data combined with cellular and biochemical studies have allowed us to propose a general, non-polymer based model for Walker-box segregation that we will test using cellular and molecular approaches. Our work on the nitrogen regulatory circuitry in B. subtilis has revealed new DNA binding modalities and a novel regulatory mechanism involving the direct enzyme of nitrogen homeostasis, glutamine synthetase. A new direction for the lab is to deduce the molecular mechanisms controlling Streptomyces development, which coincides with their production of antibiotics (secondary metabolites). Indeed, Streptomyces generate most of our current antibiotics as well as a plethora of biomedically important compounds. In the next 5 years we will expand on these efforts, but also add cellular, genetic and cryo-EM microscopy approaches to provide a more complete picture of these systems. These investigations notably intersect with the lab's interests in microbial multidrug resistance and multidrug tolerance. Indeed, while the overall goals of these studies are to determine fundamental biological principles these ongoing studies will also provide novel targets for the development of antimicrobial therapeutics, which are urgently needed given the alarming rise of multidrug resistant microbes and the scarcity of new antimicrobials in the pipeline.

抽象的 Schumacher实验室的主要重点是推断出管理基本的分子原则 涉及蛋白质核酸相互作用的生物过程。我们最近的工作已经磨练了转录 微生物中的网络和DNA分离。后者的研究发现了对分子的关键见解 使用类似肌动蛋白的和微管蛋白样NTPases分离细菌的简化系统使用的机制 质粒。但是，控制最常见的细菌分离系统的分子机制， 基于Walker-Box的系统尚不清楚，并代表了我们研究的重点。令人惊讶的是，我们的 最近表征第一个古细菌隔离系统的最新研究表明，它使用了细菌样 Walker-Box NTPase驱动DNA分离，表明Walker-Box隔离机器可能是 生物学中大多数无处不在的DNA隔离模块。这些调查还表明 古细菌种族隔离PARB蛋白具有类似于CENPA的折叠，CENPA是介导DNA的组蛋白同源物 真核生物的隔离。因此，这些研究发现了隔离中可能的进化联系 生命三个领域之间的机械。我们最近的工作提供了第一个分子观点 Walker-Box NTPases与DNA和PARB结合。这些数据与细胞和生化研究结合 允许我们提出一个通用的，非聚合物的基于沃克盒隔离的模型，我们将测试 使用细胞和分子方法。我们在枯草芽孢杆菌的氮调节电路上的工作 揭示了新的DNA结合方式和一种涉及直接酶的新型调节机制 氮稳态，谷氨酰胺合成酶。实验室的新方向是推断分子 控制链霉菌开发的机制，与其抗生素的产生相吻合 （次生代谢产物）。确实，链霉菌产生了我们目前的大多数抗生素以及大量 生物医学重要的化合物。在接下来的5年中，我们将扩大这些努力，但也会增加细胞， 遗传和冷冻EM显微镜的方法可提供这些系统的更完整的图像。这些 调查特别与实验室在微生物多药耐药性和多药的兴趣相交 宽容。确实，尽管这些研究的总体目标是确定基本的生物学原理 这些正在进行的研究还将为开发抗菌疗法的新目标提供新的目标，这 鉴于多药耐药微生物的令人震惊的上升和新的稀缺性，迫切需要迫切需要 管道中的抗菌素。