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）：在少数经过详细研究的生物体中，杆状细菌的单个环状染色体是高度组织化的，并且染色体的区域具有特定的亚细胞地址；复制起点位于旧细胞极，复制终点位于细胞分裂产生的新极附近。其他基因座在细胞中按照它们在染色体上出现的顺序排列，从起点到终点。相比之下，丝状土壤细菌天蓝色链霉菌具有大型线性基因组。人们对链霉菌孢子内线性染色体的结构知之甚少，并且在专门的气生菌丝同步分裂期间控制其分离的机制也知之甚少。这些菌丝体生物对健康和生物技术行业具有特别重要的意义，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，研究人员一直假设天蓝色链球菌的每个孢子室都有一个基因组拷贝。强有力的初步证据表明传统观点是错误的。首先，有遗传证据。参与 DNA 分离和浓缩的某些基因的突变体似乎会产生含有 DNA 的孢子，该孢子解浓缩后会显示出两个 DNA 叶。此外，可以从野生型菌株中分离出明显的遗传二倍体。其次，有细胞学证据。使用共焦激光扫描显微镜测量与野生型核仁结合的荧光染料的强度，并且该分析支持每个孢子有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有两个基因组拷贝。将探针与起始区域杂交显示出位于孢子相对两端的两个荧光焦点。此外，分区蛋白-EGFP融合体在原点附近具有结合位点，定位于成熟孢子两极的两个焦点，正如使用FISH对原点区域所观察到的那样。编码沃克型 ATP 酶的候选基因已被鉴定，该基因参与孢子内染色体位置的建立。本研究提出了完成初步遗传实验并扩展孢子基因组结构的细胞学分析的实验。编码发育相关基因组分离所需蛋白质的其他基因将通过细菌两种杂交分析来鉴定。将测试编码分离蛋白的几个已知基因的转录，以确定它们对发育调节基因的依赖性。公共健康相关性：我们希望了解 DNA 是如何在特殊的丝状生物体中分离和包装的，这种生物体产生大多数用于治疗感染和癌症的生物活性化合物。这些是我们赖以健康和生活质量的化合物的主要生产商。了解这些生物体的基本细胞功能可能会更好地生产已知和新颖的化合物，并降低购买药品的成本。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Influence of core divisome proteins on cell division in Streptomyces venezuelae ATCC 10712.

核心分裂体蛋白对委内瑞拉链霉菌 ATCC 10712 细胞分裂的影响。

DOI：
10.1099/mic.0.001015
发表时间：
2021-01-05
期刊：
Microbiology
影响因子：
1.5
作者：
Stuart Cantlay;Beer Chakra Sen;K. Flärdh;J. R. McCormick
通讯作者：
J. R. McCormick

A conserved cell division protein directly regulates FtsZ dynamics in filamentous and unicellular actinobacteria.

一种保守的细胞分裂蛋白直接调节丝状和单细胞放线菌中的 FtsZ 动力学。

DOI：
发表时间：
2021-03-17
期刊：
影响因子：
7.7
作者：
Ramos;Bush, Matthew J;Sallmen, Joseph W;Chandra, Govind;Richardson, Jake;Findlay, Kim C;McCormick, Joseph R;Schlimpert, Susan
通讯作者：
Schlimpert, Susan

A mechanism for FtsZ-independent proliferation in Streptomyces.

链霉菌中不依赖 FtsZ 的增殖机制。