RNA regulation associated with mcr11-abmR locus in M. tuberculosis

结核分枝杆菌中与 mcr11-abmR 位点相关的 RNA 调控

• 批准号: 10884585
• 负责人: Kathleen A McDonough
• 金额: $ 56.51万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10884585
• 关键词: Address; Anabolism; Bacteria; Bacterial Genes; Base Pairing; Binding; Binding Sites; Biochemical; Biological; Biological Assay; Biology; Compensation; Complex; Cryoelectron Microscopy; DNA; DNA Binding; Data; Disease; Environment; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genus Mycobacterium; Goals; Growth; Infection; Intervention; Lung; Mediating; Messenger RNA; Metabolism; Methods; Modification; Molecular; Molecular Chaperones; Mus; Mutagenesis; Mycobacterium tuberculosis; Nature; Nucleic Acid Binding; Nucleic Acids; Phase; Quantitative Reverse Transcriptase PCR; RNA; RNA Binding; RNA Recognition Motif; RNA Stability; RNA-Protein Interaction; Regulation; Regulator Genes; Reporter; Repression; Research; Resolution; Role; Structure; Structure-Activity Relationship; Testing; Transfer RNA; Tuberculosis; Untranslated RNA; Vitamin K 2; Work; fatty acid metabolism; genetic regulatory protein; genomic locus; global health; insight; novel; particle; posttranscriptional; promoter; reconstruction; response; three dimensional structure; transcription factor; transcriptome sequencing

Project summary Mycobacterium tuberculosis (Mtb) adapts to host-associated environments during infection by modulating gene expression. While transcription factors (TFs) are the most widely recognized regulators of bacterial gene expression, RNA-based factors that modulate gene expression in Mtb are less well understood. The long term goal of this project is to define fundamental mechanisms of gene regulation in Mtb, with a particular focus on the role of RNAs. The objective of this project is to define riboregulatory mechanisms associated with two products of the abmR-mcr11 gene locus in Mtb: the prototypical sRNA Mcr11 and the dual function RNA binding TF AbmR, which is required for stable mcr11 expression. We will test the hypothesis that multiple distinct regions of Mcr11 regulate gene expression by base pairing with target mRNA sequences, and characterize the impact of Mcr11 regulation in Mtb. Stable expression of Mcr11 requires AbmR, which binds ATP and contributes to central metabolism in Mtb by repressing expression of the menaquinone biosynthesis gene menE. We propose that RNA- mediated oligomerization of AbmR reduces its TF activity by converting it to a 39S complex with an alternate function. High resolution structural information regarding the nature of the RNA- AbmR complex interaction is critically needed to identify AbmR complex functions and its possible roles in Mtb biology. The specific aims of this research include: Aim 1: Define regulatory interactions of Mcr11 at the biological, molecular and genetic levels; and Aim 2: Characterize the roles of RNA-AbmR interactions at the structural and biochemical levels. These studies will combine high resolution cryo-electron microscopy with strategic mutagenesis, biochemical and biological approaches to define AbmR-RNA interactions while providing fundamental new insights into RNA-mediated regulatory mechanisms in Mtb.

项目概要 结核分枝杆菌 (Mtb) 在感染过程中通过以下方式适应宿主相关环境： 调节基因表达。虽然转录因子 (TF) 是最被广泛认可的 细菌基因表达的调节因子，调节基因表达的基于 RNA 的因子 Mtb 不太为人所知。该项目的长期目标是定义基本的 Mtb 基因调控机制，特别关注 RNA 的作用。这 该项目的目标是定义与两种产品相关的核糖调节机制 Mtb 中的 abMR-mcr11 基因位点：原型 sRNA Mcr11 和双功能 RNA 结合 TF AbmR，这是 mcr11 稳定表达所必需的。我们将检验假设 Mcr11 的多个不同区域通过与靶标碱基配对来调节基因表达 mRNA 序列，并表征 Mcr11 调节对 Mtb 的影响。稳定表达 Mcr11 需要 AbmR，它与 ATP 结合并通过以下方式促进 Mtb 的中枢代谢： 抑制甲基萘醌生物合成基因 menE 的表达。我们建议 RNA- AbmR 介导的寡聚化通过将其转化为 39S 复合物来降低其 TF 活性 替代功能。有关 RNA 性质的高分辨率结构信息 AbmR 复合体相互作用对于识别 AbmR 复合体功能及其功能至关重要 在结核分枝杆菌生物学中可能发挥的作用。本研究的具体目标包括： 目标 1：定义 Mcr11 在生物、分子和遗传水平上的调控相互作用；和目标 2： 描述 RNA-AbmR 相互作用在结构和生化水平上的作用。 这些研究将高分辨率冷冻电子显微镜与战略诱变相结合， 定义 AbmRNA-RNA 相互作用的生化和生物学方法，同时提供 对 Mtb 中 RNA 介导的调控机制的基本新见解。