Development-Associated Linear Chromosome Segregation in Streptomyces

链霉菌中发育相关的线性染色体分离

• 批准号: 10579038
• 负责人: JOSEPH R MC CORMICK
• 金额: $ 40.11万
• 依托单位: DUQUESNE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579038
• 关键词: ATP phosphohydrolase; ATPase Domain; Actinobacteria class; Alanine; Animal Model; Anthelmintics; Antibiotic Resistance; Antibiotics; Antifungal Agents; Architecture; Bacteria; Bacterial Genome; Bacterial Infections; Basic Science; Binding; Binding Proteins; Biotechnology; Biotin; Cell division; Cell physiology; Cellular biology; Cellulose; Centromere; Characteristics; Chitin; Chromosome Segregation; Chromosomes; Complex; Cytology; DNA; Deposition; Development; Ecosystem; Environment; Event; Family; Firmicutes; Gene Cluster; Gene Mutation; Genes; Genetic; Genome; Genomics; Goals; Growth; Health; Herbicides; Human; Hyphae; Immunosuppressive Agents; Knowledge; Label; Laboratories; Laboratory Study; Lead; Leucine Zippers; Life; Life Cycle Stages; Ligase; Malignant Neoplasms; Medical; Medicine; Modeling; Morphology; Motor; Nature; Nutrient; Odors; Organism; Pharmacologic Substance; Physical condensation; Plant Roots; Plants; Polymers; Production; Property; Protein Family; Protein Secretion; Proteins; Quality of life; Reproduction spores; Rod; Scanning; Side; Soil; Streptomyces; Structure; System; Time; Veterinary Medicine; anti-cancer; combat; condensin; cost; daughter cell; genetic approach; in vivo; member; mutant; next generation; novel; null mutation; protein protein interaction; segregation; yeast two hybrid system

Project Summary Streptomyces are ubiquitous filamentous sporulating soil bacteria, important in the environment because they degrade and recycle the nutrients locked in many recalcitrant natural polymers in the soil (e.g., cellulose and chitin) and produce the majority of a wide range of biologically active compounds that are used extensively in human and veterinary medicine (e.g., antibiotic, antiviral, anticancer, antitumor, antifungal, immunosuppressant and anthelmintic pharmaceuticals). Streptomyces are also well known as one of the major producers of the compounds that give soil its characteristic earthy odor. These organisms have large linear genomes. Little is known about the architecture of the linear chromosome within the Streptomyces spore and the mechanisms which govern its development-associated genome segregation and nucleoid condensation during spore formation are poorly understood. The ultimate goal from the proposed basic research is to have a better understanding of the novel cellular processes of these distinctive organisms of tremendous medical importance. The unusual mycelial life cycle and linear structure of the Streptomyces genome are likely to require some unique solutions to the fundamental problems of genome segregation and condensation. The first specific aim of the proposal is to characterize a novel small coiled-coil protein that is exclusive to this group of organisms. A genetic approach was used to provide evidence that this protein is involved in development-associated genome segregation. A proximity-labeling approach, with a promiscuous biotin ligase fusion, is being used to identify proteins that interact with this novel protein in vivo. Alanine-scanning and random PCR-directed mutageneses are being used to identify residues important for protein-protein interaction with one known binding partner. The second specific aim is to characterize type VII secretion system ATPase proteins. In addition to their functions in protein secretion, the ATPase proteins are moonlighting in development-associated segregation and nucleoid condensation. A genetic approach will be used to dissect the first type VII secretion ATPase to identify the region(s) needed for genome segregation. A genetic approach will also be used to determine the parts of both type VII secretion ATPases to identify the regions necessary for correct nucleoid condensation in the spore. Mutants lacking genes encoding both type VII secretion ATPases have genomes organized against the spore periphery (doughnut-shaped) instead of being condensed as an ellipsoid nucleoid in the center of the spore.

项目摘要 链霉菌是一种广泛存在于土壤中的丝状产孢细菌，在环境中具有重要意义 因为它们降解并回收了许多天然聚合物中的营养物质， 土壤（例如，纤维素和甲壳素），并产生大部分的广泛的生物活性 广泛用于人类和兽医学的化合物（例如，抗生素，抗病毒药， 抗癌药、抗肿瘤药、抗真菌药、免疫抑制药和驱虫药）。 链霉菌也是众所周知的化合物的主要生产者之一，使土壤 特有的泥土气味。这些生物体具有大的线性基因组。很少有人知道的 链霉菌孢子内线性染色体的结构和 控制其发育相关的基因组分离和孢子期间的类核凝聚 形成知之甚少。拟议的基础研究的最终目标是 更好地了解这些独特的生物体的新细胞过程， 医学重要性 链霉菌基因组不寻常的菌丝生命周期和线性结构可能需要 基因组分离和浓缩的基本问题的一些独特的解决方案。的 该建议的第一个具体目的是表征一种新的小卷曲螺旋蛋白，其是专属于 这群生物。遗传学方法被用来提供证据，证明这种蛋白质是 参与发育相关的基因组分离。邻近标记方法， 混杂的生物素连接酶融合，正被用来鉴定与这种新蛋白相互作用的蛋白质 in vivo.丙氨酸扫描和随机PCR定向诱变被用来鉴定残留物 对于蛋白质-蛋白质与一种已知结合配偶体的相互作用是重要的。第二个具体目标是 表征VII型分泌系统ATP酶蛋白。除了在蛋白质中的功能外， 在分泌过程中，ATP酶蛋白在发育相关的分离和类核中兼职 凝遗传学方法将用于解剖第一个VII型分泌ATP酶， 确定基因组分离所需的区域。遗传学方法也将用于 确定两种VII型分泌ATP酶的部分，以确定正确的细胞增殖所必需的区域。 孢子中的类核凝聚。缺乏编码两种VII型分泌ATP酶的基因的突变体 基因组排列在孢子外围（甜甜圈状），而不是浓缩在一起 在孢子中心呈椭圆形的类核。