Hfq RNA Chaperone and the Mechanism of RNA-Dependent Regulation

Hfq RNA 伴侣和 RNA 依赖性调节机制

• 批准号: 9353437
• 负责人: SARAH A. WOODSON
• 金额: $ 31.28万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9353437
• 关键词: Acidity; Anti-Bacterial Agents; Architecture; Arginine; Attenuated; Bacteria; Bacteria sigma factor KatF protein; Bacterial Infections; Base Pairing; Binding; Binding Sites; Biochemical; Biological Assay; Biophysics; C-terminal; Cells; Cellular Stress; Chemicals; Complementary RNA; Complex; Desiccation; Disabled Persons; Disease; Down-Regulation; Drug Targeting; Engineering; Environment; Escherichia coli; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Goals; Growth; Health; Homeostasis; Homologous Gene; Hospitals; Host Defense; Human; Human body; In Vitro; Infection; Label; Laboratories; LacZ Genes; Lead; Location; Messenger RNA; Methods; Modeling; Modification; Molecular Chaperones; Molecular Conformation; Nutrient; Outcome; Peptides; Proteins; RNA; RNA Binding; RNA Splicing; RNA annealing; RNA-Binding Proteins; RNA-Protein Interaction; Reaction; Regulation; Reporter; Repression; Research; Research Project Grants; Small RNA; Spectrum Analysis; Stress; Structure; Testing; Therapeutic; Time; Toxin; Translations; Untranslated RNA; Up-Regulation; Virulence; Virulence Factors; Work; biochemical tools; biological adaptation to stress; biophysical tools; cell growth; design; insight; mRNA Precursor; microbial disease; oxidation; pathogen; pathogenic bacteria; response; single molecule; single-molecule FRET; small molecule inhibitor; snRNP Structural Core Protein; three dimensional structure; tool

Small non-coding RNAs (sRNAs) in bacteria regulate cell growth and stress response in conjunction with an RNA chaperone protein Hfq. sRNAs control the expression of virulence factors and toxins in pathogenic bacteria, and allow bacteria to survive harsh environmental conditions such as dessication, oxidation or acidity that allow them to defeat host defenses or persist in hospital environments. Conversely, the virulence of many strains is reduced or attenuated when sRNAs are disabled. Although thousands of sRNAs have been identified in diverse bacterial species, how sRNAs and Hfq physically recognize mRNA targets to change gene expression is much less understood. The basic RNA-Hfq interaction motifs have been identified. The goal of this research is to investigate how these modular interaction motifs combine to upregulate or downregulate different gene targets. Single-molecule spectroscopy will used to study how Hfq facilitates sRNA-mRNA base pairing, while biochemical, physical and genetic methods will be used to probe the three-dimensional structure of different classes of Hfq-RNA complexes. Finally, a new model for auto-regulation of Hfq by an intrinsically disordered peptide will be investigated. Understanding how sRNAs turn genes on and off in bacteria is critical for understanding and treating microbial disease. The results of this research will uncover new antibacterial drug targets and aid the engineering of sRNAs for the control of bacterial growth. As Hfq is homologous to Sm proteins, the results will be relevant to how human Sm and Lsm proteins act in mRNA turnover and pre-mRNA splicing.

细菌中的小非编码 RNA (sRNA) 调节细胞生长和应激反应 与 RNA 伴侣蛋白 Hfq 结合。 sRNA 控制毒力的表达 抑制病原菌中的因子和毒素，使细菌能够在恶劣的环境中生存 干燥、氧化或酸性等条件使它们能够击败宿主防御或 坚持在医院环境中。相反，许多菌株的毒力降低或 当sRNA被禁用时减弱。 尽管在不同的细菌物种中已鉴定出数千种 sRNA，但 sRNA 和如何 Hfq 从物理上识别 mRNA 靶点来改变基因表达，但人们对它的了解却少之又少。 基本的 RNA-Hfq 相互作用基序已被鉴定。这项研究的目标是 研究这些模块化相互作用基序如何结合起来上调或下调 不同的基因靶标。单分子光谱将用于研究 Hfq 如何促进 sRNA-mRNA碱基配对，同时将采用生化、物理和遗传学方法 探测不同类别 Hfq-RNA 复合物的三维结构。最后，一个新的 将研究通过本质上无序的肽自动调节 Hfq 的模型。 了解 sRNA 如何在细菌中打开和关闭基因对于理解和关闭基因至关重要。 治疗微生物疾病。这项研究的结果将发现新的抗菌药物 靶向并帮助 sRNA 工程化以控制细菌生长。由于 Hfq 是 与 Sm 蛋白同源，结果将与人类 Sm 和 Lsm 蛋白的作用相关 mRNA 周转和前 mRNA 剪接。