Structural homeostasis and the assembly of a bacterial nanomachine

结构稳态和细菌纳米机器的组装

• 批准号: 9098739
• 负责人: Daniel B Kearns
• 金额: $ 30.27万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9098739
• 关键词: Animal Model; Anti-Bacterial Agents; Architecture; Area; Bacillus subtilis; Bacteria; Bacterial Infections; Behavior; Binding; Binding Sites; Biochemistry; Biology; Cell division; Cell surface; Cells; Complex; Coupled; Cytoplasm; Disease; Environment; Evolution; Face; Feedback; Filament; Flagella; Flagellin; Gammaproteobacteria; Gene Expression; Genetic; Genetic Models; Goals; Health; Homeostasis; Infection; Learning; Length; Measures; Microbial Biofilms; Microscopy; Molecular Chaperones; Mutate; Mutation; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phylogenetic Analysis; Physiology; Pilum; Post-Transcriptional Regulation; Proteins; RNA Binding; RNA-Binding Proteins; Regulation; Regulator Genes; Research; Resolution; Role; Runaway; Small RNA; Structural Genes; Structure; System; Therapeutic; Transcript; Transcriptional Regulation; Translations; Trees; Virulence; Work; cell envelope; combat; forward genetics; inhibitor/antagonist; insight; mutant; nanomachine; pathogen; pathogenic bacteria; prevent; stoichiometry; structural biology

 DESCRIPTION (provided by applicant): Bacteria build elaborate nanomachines on their cell surface to interact with their environment and cause disease. Each machine must be assembled from specific parts, in the proper order, and in precise amounts. Here we propose to study the mechanism by which a structural protein of the flagellar filament, Hag, homeostatically controls its own synthesis and governs its own assembly in Bacillus subtilis. Hag is part of a three-node negative feedback loop including FliW and the highly conserved post-transcriptional regulator of virulence gene expression, CsrA. We will determine the mechanism by which FliW antagonizes CsrA and we will determine the NMR solution structures of CsrA and FliW, separately and in complex. We will study an internal feedback loop in which the flagellin transcript helps maintain CsrA-FliW stoichiometry. We will measure the cytological parameters of filament length and elongation rate, determine how feedback regulation contributes to each parameter, and explore homeostatic regulation in another structural protein, the FlgE flagellar hook subunit. Our work will establish new paradigms for the post-transcriptional regulation of nanomachine assembly and provide new insights into the origin and evolution of virulence gene regulators. We will advance B. subtilis as a model organism for flagellar research and we use B. subtilis as a genetic platform for the expression and study of heterologous regulators from distantly related pathogenic bacteria. Finally, we have isolated drugs that antagonize CsrA when added to whole cells will characterize the anti-CsrA drugs as candidate antibacterial therapeutics.

 描述（由适用提供）：细菌在其细胞表面上建立精美的纳米机器，以与环境相互作用并引起疾病。必须按适当的顺序和精确的数量从特定零件组装每台机器。在这里，我们建议研究鞭毛细丝的结构蛋白，hag，稳态控制其自身的合成，并控制其在枯草芽孢杆菌中的组装。巫婆是三节点负反馈循环的一部分，包括FLIW和高度配置的病毒基因表达的转录后调节剂CSRA。我们将确定FLIW拮抗CSRA的机制，并将分别和复杂的CSRA和FLIW的NMR溶液结构确定。我们将研究一个内部反馈回路，其中鞭毛蛋白转录本有助于维持CSRA-FLIW化学计量。我们将测量细胞长度和伸长率的细胞学参数，确定反馈调节如何有助于每个参数，并探索另一种结构蛋白（Flge Flagllar钩子亚基）中的稳态调节。我们的工作将为纳米机械组装的转录后调节建立新的范式，并为病毒基因调节剂的起源和进化提供新的见解。我们将促进枯草芽孢杆菌作为旗杆研究的模型生物，并将枯草芽孢杆菌作为一种遗传平台，用于表达和研究来自明显相关的致病细菌的异源调节剂。最后，当将抗CSRA药物添加到整个细胞中时，我们有分离的药物可以拮抗CSRA，将抗CSRA药物作为候选抗菌治疗。