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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的舒马赫实验室的中心重点是推论控制基本原理的分子原理涉及蛋白质-核酸相互作用的生物过程。我们最近的工作重点是转录微生物中的网络和 DNA 分离。后面的研究揭示了分子生物学的关键见解简化系统利用肌动蛋白样和微管蛋白样 NTPase 来分离细菌的机制质粒。然而，控制最常见细菌分离系统的分子机制，基于 Walker-box 的系统仍然不清楚，也是我们研究的一个主要焦点。引人注目的是，我们的最近的研究描述了第一个古菌分离系统的特征，表明它利用了一种类似细菌的 Walker-box NTPase 驱动 DNA 分离，表明 Walker-box 分离机器可能是生物学中最普遍的 DNA 分离模块类型。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 的组蛋白同源物真核生物中的分离。因此，这些研究揭示了隔离中可能的进化联系生活三个领域之间的机器。我们最近的工作提供了第一个分子观点 Walker-box NTPase 与 DNA 和 ParB 结合。这些数据与细胞和生化研究相结合使我们能够提出一个通用的、基于非聚合物的 Walker-box 分离模型，我们将对其进行测试使用细胞和分子方法。我们在枯草芽孢杆菌氮调节回路方面的工作已经揭示了新的 DNA 结合方式和涉及直接酶的新调控机制氮稳态，谷氨酰胺合成酶。实验室的一个新方向是推论分子控制链霉菌发育的机制，这与它们生产抗生素是一致的（次级代谢产物）。事实上，链霉菌产生了我们目前使用的大部分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，同时还会增加蜂窝、遗传和冷冻电镜显微镜方法可以提供这些系统的更完整的图像。这些研究与实验室对微生物多药耐药性和多药性的兴趣密切相关宽容。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目标，鉴于多重耐药微生物的惊人增加和新药的稀缺，迫切需要管道中的抗菌剂。