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CHROMOSOME SEGREGATION IN A FILAMENTOUS BACTERIUM

丝状细菌中的染色体分离

基本信息

批准号：
8036672
负责人：
JOSEPH R MC CORMICK
金额：
$ 26.94万
依托单位：
DUQUESNE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8036672
关键词：
ATP phosphohydrolase Actinobacteria class Address Animal Model Anthelmintics Antibiotic Therapy Antifungal Antibiotics Architecture Attention Bacteria Bacterial Proteins Binding Binding Sites Biotechnology Candidate Disease Gene Cell Wall Cell division Cell physiology Cells Cellular biology Chromosome Positioning Chromosome Segregation Chromosomes Complex DNA DNA analysis DNA biosynthesis Dependence Development Developmental Cell Biology Developmental Gene Diploidy Fermentation Fluorescent Dyes Fluorescent in Situ Hybridization Gene Expression Regulation Genes Genetic Genetic Transcription Genome Glean Growth Health Herbicides Hyphae Immunosuppressive Agents Industry Infection Knowledge Laboratories Laser Scanning Confocal Microscopy Lead Life Cycle Stages Lobe Location Malignant Neoplasms Measures Medical Medicine Microbiology Molds Mycobacterium tuberculosis Nature Organism Outcome Physical condensation Ploidies Process Production Prokaryotic Cells Proteins Quality of life Regulation Regulator Genes Relative (related person)Replication Origin Reproduction spores Research Research Personnel Research Project Grants Shapes Site Soil Solutions Streptomyces Streptomyces coelicolor Streptomyces coelicolor A3(2)System Testing United States Walkers antitumor drug cost daughter cell design enhanced green fluorescent protein interest microorganism mutant novel research study retinal rods segregation yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): In the few organisms where it has been studied in detail, the single circular chromosome of a rod-shaped bacterium is highly organized and regions of the chromosome have particular subcellular addresses; origins of replication are located at the old cell poles, replication termini are near the new poles derived from cell division. Other loci are arranged in the cell in the order that they appear on the chromosome, origin to terminus. In contrast, the filamentous soil-dwelling bacterium Streptomyces coelicolor has a large linear genome. Little is known about the architecture of the linear chromosome within the Streptomyces spore and the mechanisms which govern its segregation during synchronous division of specialized aerial hyphae are poorly understood. These mycelial organisms are of particular and vital importance to health and biotechnology industries because they produce many biologically active compounds such as antibiotics, antifungals, antihelminthics, herbicides and antitumor drugs. For years researchers have assumed that there is a single copy of the genome per spore compartment of S. coelicolor. Strong preliminary evidence suggests that conventional wisdom is incorrect. First, there is genetic evidence. Mutants of certain genes involved in DNA segregation and condensation appear to produce spores containing DNA that is decondensed showing two lobes of DNA. In addition, apparent genetic diploids can be isolated from the wild type strain. Second, there is cytological evidence. Confocal laser scanning microscopy was used to measure the intensity of fluorescent dye bound to the wild type nucleoid, and the analysis supports the hypothesis of two genome copies per spore. Similarly, preliminary analysis of fluorescent in situ hybridization (FISH) results point to two copies of the genome per wild type spore. Hybridizing a probe to the origin region shows two fluorescent foci located at opposite ends of the spore. Furthermore, a partition protein- EGFP fusion, with binding sites near the origin, localizes in two foci to the poles of maturing spores, as was observed for the origin region using FISH. A candidate gene, encoding a Walker-type ATPase, involved in the establishment of the chromosome position within the spore has been identified. This study proposes experiments to complete the preliminary genetic experiments and extend the cytological analysis of the genome architecture in the spore. Additional genes encoding proteins required for development-associated genome segregation will be identified by bacterial two hybrid analyses. Transcription of several known genes encoding segregation proteins will be tested for their dependence on developmental regulatory genes. PUBLIC HEALTH RELEVANCE: We wish to understand how DNA is segregated and packaged in the special filamentous organisms that produce the majority of biologically active compounds used to treat infection and cancer. These are major producers of the compounds we all rely on for our health and quality of life. Basic knowledge of fundamental cell functions of these organisms may lead to better production of known and novel compounds and lower the cost of buying medicines.

描述（由申请人提供）：在对其进行详细研究的少数生物体中，杆状细菌的单个环状染色体高度组织化，染色体区域具有特定的亚细胞地址;复制起点位于旧细胞极点，复制末端靠近细胞分裂产生的新极点。其他基因座在细胞中的排列顺序是它们在染色体上出现的顺序，从起点到末端。相比之下，丝状土壤细菌天蓝色链霉菌具有大的线性基因组。关于链霉菌孢子内线性染色体的结构知之甚少，并且在特化气生菌丝的同步分裂期间控制其分离的机制知之甚少。这些菌丝体生物体对健康和生物技术工业具有特别和至关重要的意义，因为它们产生许多生物活性化合物，例如抗生素、抗真菌剂、抗蠕虫剂、除草剂和抗肿瘤药物。多年来，研究人员一直认为，有一个单拷贝的基因组每个孢子室的S。天蓝色。强有力的初步证据表明，传统智慧是不正确的。首先，有遗传学证据。参与DNA分离和凝聚的某些基因的突变体似乎产生含有DNA的孢子，该孢子解凝聚，显示出两个DNA叶。此外，可以从野生型菌株中分离出明显的遗传二倍体。第二，有细胞学证据。共聚焦激光扫描显微镜用于测量结合到野生型类核的荧光染料的强度，并且分析支持每个孢子两个基因组拷贝的假设。同样，荧光原位杂交（FISH）结果的初步分析表明，每个野生型孢子的基因组有两个拷贝。将探针杂交到起始区域显示位于孢子相对端的两个荧光焦点。此外，分区蛋白- EGFP融合，与结合位点附近的起源，定位在两个焦点的两极成熟孢子，所观察到的起源区域使用FISH。一个候选基因，编码步行型ATP酶，参与建立孢子内的染色体位置已被确定。这项研究提出了实验，以完成初步的遗传实验，并扩大孢子中的基因组结构的细胞学分析。将通过细菌双杂交分析鉴定编码发育相关基因组分离所需蛋白质的其他基因。几个已知的基因编码分离蛋白的转录将测试其依赖于发育调控基因。公共卫生相关性：我们希望了解DNA是如何在特殊的丝状生物中分离和包装的，这些丝状生物产生了用于治疗感染和癌症的大多数生物活性化合物。这些是我们都依赖于我们的健康和生活质量的化合物的主要生产商。对这些生物体的基本细胞功能的基本知识可能会导致更好地生产已知和新的化合物，并降低购买药物的成本。