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: 10543420
• 负责人: Maria Schumacher
• 金额: $ 38.8万
• 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 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10543420
• 关键词: Actins; Antibiotics; Bacillus subtilis; Binding; 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; Work; antimicrobial; combat; drug resistant microorganism; insight; interest; microbial; multidrug tolerance; novel; nucleoside triphosphatase; rational design; secondary metabolite; 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.

摘要 舒马赫实验室的中心焦点是推导出支配基本的分子原理， 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在转录 网络和微生物中的DNA分离。后一项研究揭示了分子生物学的关键见解， 采用肌动蛋白样和微管蛋白样NTPases分离细菌的简化系统所利用的机制 质粒。然而，控制最常见的细菌分离系统的分子机制， 基于Walker-box的系统仍然不清楚，并且代表了我们研究的主要焦点。引人注目的是，我们的 最近的研究表明，第一个古细菌分离系统利用了一种类似细菌的 Walker-box NTT驱动DNA分离，表明Walker-box分离机制可能是DNA分离的主要机制。 生物学中最普遍的DNA分离模块。这些调查还显示， 古细菌分离ParB蛋白具有与CenpA相似的折叠，CenpA是介导DNA的组蛋白同系物 真核生物中的分离因此，这些研究揭示了种族隔离中可能的进化联系 在生命的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， 与DNA和ParB结合的Walker-box NTP。这些数据与细胞和生物化学研究相结合 使我们能够提出一个通用的，非聚合物为基础的模型沃克盒隔离，我们将测试 使用细胞和分子方法。我们对B中氮调节回路的研究。枯草杆菌有 揭示了新的DNA结合方式和一种新的调节机制，涉及直接酶， 氮平衡谷氨酰胺合成酶实验室的一个新方向是推导出 控制链霉菌发育的机制，这与它们生产抗生素的过程一致 （次级代谢物）。事实上，链霉菌产生了我们目前的大多数抗生素以及大量的抗生素。 生物医学上重要的化合物。在接下来的5年里，我们将扩大这些努力，但也增加蜂窝， 遗传学和冷冻EM显微镜方法，以提供这些系统的更完整的图片。这些 研究与实验室在微生物多药耐药性和多药耐药方面的兴趣有明显的交叉。 宽容事实上，虽然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抗菌疗法的开发提供新的靶点， 鉴于多药耐药微生物的惊人增长和新药物的缺乏， 抗菌药物在管道中。