Transcriptome and proteome remodeling by Mycobacterium tuberculosis MazF toxins

结核分枝杆菌 MazF 毒素的转录组和蛋白质组重塑

基本信息

批准号：
9886870
负责人：
NANCY ANN WOYCHIK
金额：
$ 62.63万
依托单位：
RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-12-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9886870
关键词：
AIDS/HIV problem Bacteria Bioinformatics Biological Biology Biotin Cause of Death Cells Characteristics Chemicals Chemistry Computational Biology Consensus Sequence Data Detection Disease Endoribonucleases Exhibits Exposure to Family Family member Genes Genetic Goals Growth HIV Harvest Human Immune Immune response Immune system Individual Knowledge Laboratories Lead M. tuberculosis genome Mass Spectrum Analysis Mediating Messenger RNA Metabolic Molecular Mycobacterium tuberculosis National Institute of Allergy and Infectious Disease Operon Organism Physiologic Monitoring Physiological Physiology Protein Biosynthesis Proteins Proteome RNA Research Ribosomal RNA Role Signal Transduction Site Specificity Strategic Planning Stress System Technology Testing Therapeutic Time Toxin Transfer RNA Tuberculosis Validation Work alpha Toxin antimicrobial antitoxin assault cell growth design genome-wide improved insight latent infection macrophage member metabolic rate multidisciplinary novel therapeutics prevent protein complex response tool transcriptome transcriptome sequencing

项目摘要

Project Summary Mycobacterium tuberculosis (Mtb) has adapted to survive a wide range of assaults—from our immune response to antimicrobial therapeutics—intended to eradicate the organism. However, the molecular switches that enable Mtb to endure these stresses, to slow replication or to become dormant as a latent tuberculosis (TB) infection are not known. Emerging studies on the molecular underpinnings of stress survival generally point to a major role for toxin-antitoxin (TA) systems, which are operons comprising adjacent genes encoding two small proteins, a toxin and its cognate antitoxin that inhibits toxin activity in the TA protein-protein complex. However, several bottlenecks have impeded progress toward rigorous testing of this provocative association. This proposal enlists a strong multidisciplinary team with expertise in all core components of the proposed work—RNA-seq, TA systems, Mtb biology/physiology and bioinformatics/computational biology. The genome-scale approach developed in the PI’s laboratory, 5’ RNA-seq, will be used to overcome these obstacles as they apply to the eleven-member MazE (antitoxin) – MazF (toxin) family in Mtb. 5’ RNA-seq will facilitate comprehensive detection of MazF targets in the Mtb transcriptome under unstressed conditions or after exposure to stresses that are relevant to latent TB infection. Finally, the impact of MazF toxins on the Mtb proteome will be investigated. Collectively, these approaches will identify the environmental signals that trigger toxin activation in Mtb, provide an accurate snapshot of RNAs targeted by MazF toxins under these metabolic states, and reveal clues to how toxin-mediated RNA cleavage alters Mtb physiology. These goals align well with “Priority 1: Improve Fundamental Knowledge of TB” of the five components of the NIAID Strategic Plan for Tuberculosis Research released in September 2018.

项目概要结核分枝杆菌 (Mtb) 已经适应了我们免疫反应的多种攻击抗菌疗法——旨在根除微生物，然而，分子开关能够实现这一点。结核分枝杆菌承受这些压力，减缓复制或作为潜伏性结核病 (TB) 感染进入休眠状态目前尚不清楚压力生存的分子基础。毒素-抗毒素（TA）系统的作用，该系统是由编码两个小蛋白质的相邻基因组成的操纵子，一种毒素及其同源抗毒素，可抑制 TA 蛋白-蛋白复合物中的毒素活性。瓶颈阻碍了对这一挑衅性协会进行严格测试的进展。强大的多学科团队，在拟议工作的所有核心组成部分（RNA-seq、TA）方面拥有专业知识系统、结核分枝杆菌生物学/生理学和生物信息学/计算生物学基因组规模方法。 PI 实验室开发的 5’RNA-seq 将用于克服这些障碍，因为它们适用于 Mtb 中的 11 名 MazE（抗毒素）- MazF（毒素）家族将促进全面检测。在无应激条件下或暴露于以下应激后，Mtb 转录组中的 MazF 靶标的数量最后，将研究 MazF 毒素对 Mtb 蛋白质组的影响。总的来说，这些方法将识别触发结核分枝杆菌毒素激活的环境信号，提供在这些代谢状态下 MazF 毒素靶向的 RNA 的准确快照，并揭示如何毒素介导的 RNA 裂解改变 Mtb 生理学，这些目标与“优先事项 1：改善”非常一致。 NIAID 结核病研究战略计划五个组成部分的“结核病基础知识” 2018 年 9 月发布。