Genome exploration through toxin-mediated ribosome stalling

通过毒素介导的核糖体停滞进行基因组探索

基本信息

批准号：
10396107
负责人：
NANCY ANN WOYCHIK
金额：
$ 38.38万
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：
2020
资助国家：
美国
起止时间：
2020-05-07 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10396107
关键词：
Antibiotics Bacteria Biological Cells Characteristics Chemicals Codon Nucleotides Computer Analysis Consensus Sequence Data Data Set Development Diagnostic Disease Endoribonucleases Event Exhibits Family member Genetic Genome Growth HIV Harvest Human Immune Immune response Immune system Individual Infection M. tuberculosis genome Maps Mass Spectrum Analysis Mediating Messenger RNA Methods Molecular Mycobacterium tuberculosis Open Reading Frames Operative Surgical Procedures Pathogenesis Pathway interactions Peptides Persons Physiology Positioning Attribute Process Proteome Proteomics RNA Research Resources Ribosomal Proteins Ribosomal RNA Ribosomes Risk Sampling Series Site Specificity Stress System Technology Therapeutic Time Toxin Transcript Transfer RNA Treatment Protocols Tuberculosis annotation system antimicrobial antitoxin assault base cell growth cellular targeting effective therapy genome annotation genome-wide improved latent infection novel pathogen response rib bone structure ribosome profiling screening tool tool development transcriptome sequencing

项目摘要

Project Summary Approximately 90% of individuals infected with Mycobacterium tuberculosis (Mtb) develop an asymptomatic latent infection, which is non-infectious. Although some individuals may eradicate this infection, those who do not comprise a large reservoir of persons who can convert from latent to active TB, which is infectious. Reactivation is especially likely in immune-compromised individuals, including those infected with HIV. The molecular switches that enable Mtb to slow or stop replication, become dormant and establish latent TB infection are poorly characterized. A thorough understanding of these switches is critical for development of 1) diagnostics to enable prediction of reactivation risk and 2) shorter, more effective treatment regimens for latent TB infection. Toxin-antitoxin (TA) systems are strongly implicated in establishment of latent TB infection because their toxin components typically downregulate Mtb cell growth and are activated in response to stresses relevant to this state. Yet, the extraordinary redundancy of TA systems make determination of the individual contributions of each toxin challenging using conventional genetic and molecular biological approaches. We propose to use a powerful battery of genome-scale tools to track the fate of transcripts, ribosomes and proteins in response to activation of a subset of tRNA-cleaving toxins to understand the molecular mechanisms that underlie stress survival. We then exploit our finding that the codon-specific ribosome-stalling characteristic of these toxins identifies novel ORFs and apply this as a reliable tool for improved Mtb genome annotation.

项目概要大约 90% 的结核分枝杆菌 (Mtb) 感染者会出现无症状症状潜伏感染，即无传染性。尽管有些人可以根除这种感染，但那些做到了的人不包括大量可以从潜伏性结核病转变为具有传染性的活动性结核病的人群。免疫功能低下的个体尤其容易重新激活，包括感染艾滋病毒的人。这分子开关使 Mtb 减慢或停止复制、进入休眠状态并建立潜伏性 TB 感染的特征很差。彻底了解这些开关对于 1) 诊断的开发至关重要能够预测再激活风险；2）针对潜伏性结核感染提供更短、更有效的治疗方案。毒素-抗毒素（TA）系统与潜伏性结核感染的建立密切相关，因为它们的毒素成分通常会下调结核分枝杆菌细胞的生长，并响应与此相关的压力而被激活状态。然而，TA系统的巨大冗余决定了个体的贡献每种毒素都具有挑战性，无法使用传统的遗传和分子生物学方法。我们建议使用一个强大的基因组规模工具组，用于跟踪转录本、核糖体和蛋白质的命运，以响应激活一部分 tRNA 裂解毒素，以了解压力背后的分子机制生存。然后，我们利用我们的发现，即这些毒素的密码子特异性核糖体停滞特征识别新的 ORF 并将其用作改进 Mtb 基因组注释的可靠工具。