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： 所有生物具有共同的遗传密码。从哺乳动物细胞的核到 最小的细菌，我们都使用DNA编码驱动生命多样性的性状和蜂窝过程 在地球上。在此过程中，传递基因组是遗传力和错误的最基本形式 可能会产生后果。因此，所有生物都会进行可靠的复制非常重要 和细胞分裂过程中DNA的分离，并且生物体保持保护核材料的方法 如果发生细胞应力。为了研究DNA结构的形式和功能，我提出了利用 冷冻电子断层扫描（Cryo-ET）方法的最新进展，可视化细菌 核苷，作为理解体内染色体结构的模型，并开发工具和最佳 研究细胞最重要的分子的实践。使用冷冻电子的资源 UW-Madison的显微镜研究中心，模型生物的冷冻-ET Rhodobacter Sphaeroides将会 允许研究体内核结构。 Rhodobacter sphaeroides是研究 核苷，不仅是因为其大小与冷冻-ET相吻合，还因为其两个染色体需要 唯一的复制动力学。这项工作旨在观察核苷的结构和相互作用 Sphaeroides河的细胞周期。结果具有强大的潜力来提供翻译中断 通过并将建立分子工具和方法，这些工具和方法将超出细菌学之外的相关性。 此外，这项工作将研究身体应激和 已经显示了抗生素处理，初步数据。重要的是，研究核的压实 预计抗生素应力后的材料将产生新的知识，从而更好地理解 细菌在抗生素治疗过程中使用的机制生存和持续。潜在影响研究 关于抗生素耐药性。