Form and function of the bacterial nucleoid: structural studies using cryo-ET

细菌核的形式和功能：使用冷冻电子断层扫描进行结构研究

• 批准号: 10534657
• 负责人: Daniel Parrell
• 金额: $ 6.98万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10534657
• 关键词: Anatomy; Animal Model; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Bacteria; Bacterial Infections; Bacteriology; Butyrates; Cell Cycle; Cell Division Process; Cell Nucleus; Cell division; Cell physiology; Cells; Cellular Stress; Cellular Structures; Cellular biology; Characteristics; Chromosome Structures; Chromosomes; Crowding; Cryo-electron tomography; Cryoelectron Microscopy; Cytoplasmic Granules; Cytoplasmic Structures; DNA; DNA Packaging; DNA Structure; DNA biosynthesis; Data; Diffuse; Electroconvulsive Therapy; Environment; Event; Exhibits; Fluorescence Microscopy; Genes; Genetic; Genetic Code; Genome; Health; Heritability; Human; Knowledge; Label; Life; Mammalian Cell; Membrane; Methodology; Methods; Modeling; Molecular; Nuclear; Nuclear Structure; Nutrient; Organism; Phenotype; Physical condensation; Physiological; Physiological Processes; Planet Earth; Positioning Attribute; Process; Research; Resolution; Resources; Rhodobacter sphaeroides; Ribosomes; Sampling; Slice; Stains; Stress; Structure; System; Temperature; Thick; Tomogram; Work; daughter cell; genetic information; in vivo; mutant; response; segregation; structural biology; tool; trait

Project Summary/Abstract: All organisms share a common genetic code. From the nucleus of mammalian cells to the nucleoid of the smallest bacterium, we all use DNA to encode the traits and cellular processes that drive the diversity of life on earth. Passing along one’s genome is the most fundamental form of heritability and errors in this process can have dire consequences. It is therefore of great importance that all organisms carry out reliable replication and segregation of DNA during cell division, and that organisms maintain methods to protect nuclear material in the event of cellular stress. To study the form and function of DNA structures, I propose taking advantage of the recent advancements in cryoelectron tomography (cryo-ET) methodology to visualize the bacterial nucleoid, as a model for understanding in vivo chromosome structure and to develop the tools and best practices for the study of the cells most important molecule. Using the resources of the Cryoelectron Microscopy Research Center at UW-Madison, cryo-ET of the model organism Rhodobacter sphaeroides will allow the study of in vivo nuclear structures. Rhodobacter sphaeroides is an ideal organism for studying the nucleoid, not only because its size is amenable to cryo-ET, but also because its two chromosomes require unique replication dynamics. This work seeks to observe the structure and interactions of the nucleoid during the cell cycle of R. sphaeroides. The results of which have a strong potential to provide translational break throughs and will build up molecular tools and methods that will have relevance beyond bacteriology. Additionally, this work will study the compaction of the bacterial nucleoid during physiological stress and antibiotic treatments, preliminary data have been shown. Importantly, studying the compaction of nuclear material following antibiotic stress is expected to generate new knowledge leading to a better understanding of the mechanisms bacteria use to survive and persist during antibiotic treatment. Potentially impacting the study on antibiotic resistance.

项目摘要/摘要： 所有生物体都有共同的遗传密码，从哺乳动物细胞的细胞核到细胞核。 作为最小的细菌，我们都使用 DNA 来编码驱动生命多样性的特征和细胞过程 在地球上传递一个人的基因组是遗传性和这一过程中的错误的最基本形式。 因此，所有生物体进行可靠的复制非常重要。 和细胞分裂过程中 DNA 的分离，以及生物体维持保护核材料的方法 为了研究 DNA 结构的形式和功能，我建议利用 冷冻电子断层扫描（cryo-ET）方法的最新进展使细菌可视化 nucleoid，作为理解体内染色体结构并开发工具和最佳模型的模型 利用冷冻电子资源研究细胞最重要分子的实践。 威斯康辛大学麦迪逊分校显微镜研究中心，模型生物类球红杆菌的冷冻电子断层扫描将 允许研究体内核结构。球形红细菌是研究细胞核结构的理想生物体。 nucleoid，不仅因为它的大小适合冷冻电子断层扫描，还因为它的两条染色体需要 这项工作旨在观察核素复制过程中的结构和相互作用。 R. sphaeroides 的细胞周期，其结果具有提供翻译中断的强大潜力。 通过并将建立具有超越细菌学意义的分子工具和方法。 此外，这项工作将研究生理应激期间细菌核的压缩和 抗生素治疗，初步数据已显示，重要的是，研究核的压实。 抗生素应激后的材料预计会产生新的知识，从而更好地理解 细菌在抗生素治疗期间生存和持续存在的机制可能会影响研究。 关于抗生素耐药性。