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：
10307528
负责人：
NANCY ANN WOYCHIK
金额：
$ 60.96万
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 2022-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10307528
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.

项目摘要结核分枝杆菌（MTB）已经适应了广泛的攻击，从我们的免疫冲突中对于抗菌理论 - 旨在放射性生物体。但是，启用的分子开关 MTB忍受这些应力，缓慢复制或作为潜在结核病（TB）感染而休眠不知道。关于应力存活的分子基础的新兴研究通常指向主要毒素 - 抗毒素（TA）系统的作用，这些系统是完成编码两个小蛋白的相邻基因的歌剧， A毒素及其同源抗毒素抑制TA蛋白蛋白质复合物中毒素活性。但是，有几个瓶颈阻碍了对这种挑衅性关联进行严格测试的进展。该提议征集了一个强大的多学科团队系统，MTB生物学/生理学和生物信息学/计算生物学。基因组级方法在Pi的实验室中开发的5'RNA-Seq将用于克服这些障碍，因为它们适用于十一人民的迷宫（抗毒素） - MTB中的MAZF（毒素）家族。 5'RNA-seq将有助于全面检测 MTB转录组中的MAZF靶标在无重理条件下或暴露于应力之后与潜在结核病感染有关。最后，将研究MAZF毒素对MTB蛋白质组的影响。总的来说，这些方法将确定在MTB中触发毒素激活的环境信号，提供在这些代谢状态下，MAZF毒素针对的RNA的准确快照，并揭示了如何毒素介导的RNA裂解改变了MTB生理学。这些目标与“优先1：改进”很好地吻合结核病研究的NIAID战略计划的五个组成部分的结核病知识 2018年9月发行。