Control of Bacillus Anthracis Spore Formation

炭疽芽孢杆菌孢子形成的控制

• 批准号: 7652115
• 负责人: Philip C Hanna
• 金额: $ 34.08万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-15 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7652115
• 关键词: Alveolar; Amino Acids; Anthrax disease; Antibiotic Therapy; Antibiotics; Bacillus (bacterium); Bacillus anthracis; Bacillus anthracis spore; Bacterial Spores; Binding; Biological Assay; Chemistry, Other; Class; Databases; Dose; Event; Extracellular Domain; Family; Family member; Germination; Infection; Integration Host Factors; Intervention; Knowledge; Libraries; Lung; Lymphatic; Measurement; Medical; Membrane; Operon; Pattern; Peptides; Phage Display; Phagocytes; Phagocytosis; Pharmaceutical Preparations; Preclinical Drug Evaluation; Process; Proteins; Range; Reproduction spores; Research; Risk; Role; Screening procedure; Signal Transduction; Structure; Supervision; System; Testing; Time; Transmembrane Domain; Treatment Protocols; Yeasts; analog; assay development; base; day; drug intolerance; extracellular; gene function; genetic analysis; inhibitor/antagonist; mutant; novel; nucleoside analog; pathogen; protein protein interaction; protein structure function; receptor; small molecule; tool; uptake; yeast two hybrid system

Spore germination represents a pivotal intervention point for controlling anthrax infection. Germination occurs after uptake by alveolar phagocytes, concurrent with their transport to the lymphatics. However, the timing of the initial association of spores with phagocytes varies depending on inoculum's dose and host factors. Spores are phagocytosed soon after exposure or can linger in the lung up to 100 days before initiating a lethal infection. Spores are hardy, not easily cleared from the body and not readily susceptible to antibiotics; this necessitates prolonged antibiotic treatment & real risks of drug intolerances. Abilities to block or delay spore germination could be utilized to block the infectious cycle. Alternatively, efficiently inducing germination, under controlled medical supervision, may render the bacillus vulnerable to antibiotics, shortening the requirement for lengthy antibiotic regimens. Aim 1" Develop 96-well platforms for screening compounds that act to either block or stimulate germination of anthrax spores and screening for stimulators and inhibitors of germination. Development of these assays will be of use to other research groups, and will facilitate the mechanistic studies of the other aims. Aim 2" GerA-like protein structure/function analysis will determine the membrane topology of the 6 gerA family of B. anthracis operons, determine protein-protein interactions between the 3 GerA-like proteins encoded by each tricistronic operon and examine potential physical interactions ("cross-talk") between the proteins of the 6 family members. Aim 3: Determine the spores' repertoire of non-gerA-like proteins that form the complete germination machinery using yeast two-hybrid analysis, phage display, and genetic analysis of hyper germination mutants. Knowledge of the spore proteins responsible for germination, as well as their functions and extracellular domains will directly test the hypothesis that the rapid & dramatic switch from dormancy to growing pathogen is controlled by an intricate membrane sensing/signaling system. It will also facilitate discovery of compounds that can be used to control these germination events.

孢子萌发是控制炭疽感染的关键干预点。 萌发发生在被肺泡吞噬细胞摄取后，同时它们被运输到 反物质。然而，孢子与吞噬细胞最初结合的时间不同 这取决于接种物的剂量和宿主因素。孢子在接触后很快被吞噬， 可在肺部滞留长达100天，然后引发致命感染。孢子是哈代，不容易 从体内清除并且不容易对抗生素敏感;这需要长时间 抗生素治疗和药物不耐受的真实的风险。阻止或延迟孢子萌发的能力 可以用来阻断传染循环或者，有效诱导发芽，在 控制医疗监督，可能使杆菌容易受到抗生素，缩短 需要长期的抗生素治疗方案。目标1”开发用于筛选的96孔平台 用于阻断或刺激炭疽孢子萌发的化合物， 发芽的刺激剂和抑制剂。这些检测方法的发展将有助于其他 研究小组，并将促进其他目标的机制研究。Aim 2”类GerA 蛋白质结构/功能分析将确定B的6 gerA家族的膜拓扑结构。 炭疽操纵子，决定蛋白质之间的相互作用3 GerA样蛋白编码 每个三顺反子操纵子，并检查潜在的物理相互作用（“串扰”）之间的 6个家族成员的蛋白质。目的3：确定非gerA样蛋白的孢子库 使用酵母双杂交分析，噬菌体 超发芽突变体的展示和遗传分析。孢子蛋白的知识 负责发芽，以及他们的功能和胞外结构域将直接测试 假设从休眠到生长的病原体的快速和戏剧性的转变是由一个 复杂的膜传感/信号系统。它还将促进发现化合物， 来控制这些发芽事件。