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

基本信息

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
项目类别：
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

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

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项目摘要

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.

项目摘要结核分枝杆菌（MTB）适应与宿主相关的环境，在感染期间通过调节基因表达。而转录因子（TFS）是最广泛认可的细菌基因表达的调节剂，基于RNA的因子，这些因素调节基因表达 MTB不太了解。该项目的长期目标是定义基本 MTB中基因调节的机制，特别关注RNA的作用。这该项目的目的是定义与两种产品相关的核糖调节机制 MTB中的ABMR-MCR11基因基因座：原型SRNA MCR11和双函数RNA 结合TF ABMR，这是稳定的MCR11表达所必需的。我们将检验假设 MCR11的多个不同区域通过基础配对来调节基因表达 mRNA序列，并表征MCR11调节在MTB中的影响。稳定的表达 MCR11需要ABMR，该ABMR结合ATP并促进MTB中的中央代谢甲苯丙胺生物合成基因的表达。我们提出RNA- 介导的ABMR的寡聚化通过将其转换为39S复合物与替代功能。有关RNA的性质的高分辨率结构信息 ABMR复合物相互作用至关重要，以识别ABMR复合功能及其ITS MTB生物学的可能作用。这项研究的具体目的包括：目标1：定义 MCR11在生物学，分子和遗传水平上的调节相互作用；和目标2：表征RNA-ABMR相互作用在结构和生化水平上的作用。这些研究将结合高分辨率的冷冻电子显微镜与策略诱变，在提供的同时定义ABMR-RNA相互作用的生化和生物学方法对MTB中RNA介导的调节机制的基本新见解。