Mechanisms of ESX-1-dependent Gene Expression in Pathogenic Mycobacteria

病原分枝杆菌中 ESX-1 依赖性基因表达的机制

基本信息

批准号：
10078256
负责人：
Patricia A Champion
金额：
$ 19.31万
依托单位：
UNIVERSITY OF NOTRE DAME
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：
2020-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10078256
关键词：
Address Affinity Bacteria Biochemical Biological Cell membrane Cytolysis Cytoplasm DNA Data Diagnosis ELF3 gene Epidemic Fishes Gene Expression Gene Expression Regulation Genes Genetic Genetic Screening Genetic Transcription Genus Mycobacterium Goals Human Infection Integral Membrane Protein Knowledge Laboratories Lead Left Link Mediating Membrane Metabolism Mission Modeling Molecular Mycobacterium marinum Mycobacterium tuberculosis Outcome Pathogenesis Pathogenicity Pathway interactions Phagosomes Phase Prevention Proteins Public Health Publishing Regulation Research Role Signal Pathway Signal Transduction Signal Transduction Pathway Solid System Testing Tuberculosis United States National Institutes of Health Virulence Factors Work gene discovery genetic testing innovation insight macrophage mycobacterial pathogen prevent promoter response therapeutic development transcription factor tuberculosis treatment vaccine development

项目摘要

PROJECT SUMMARY: The applicant’s laboratory discovered that the assembly of the ESX-1 translocon in the cytoplasmic membrane regulates gene expression. The long-term goal of the applicant is to understand the biological significance of ESX-1-dependent changes in gene expression. The applicant proposes that the assembly of the ESX-1 trans- locon elicits gene expression pathways that couple phagosomal lysis and adaptation to cytoplasmic exposure. The overall objective of this proposal is to explore how the ESX-1 translocon regulates gene expression, which will contribute to addressing the applicant’s long-term goal. The central hypothesis is that there is an uncharac- terized transcription response network connecting the ESX-1 translocon to widespread changes in gene expres- sion, potentially linking phagosomal lysis and cytoplasmic adaptation. The rationale is that the successful com- pletion of the proposed work will provide insight into the fundamental mechanisms linking lysis of the phagosomal membrane by ESX-1 with changes in gene expression. The central hypothesis will be tested by following these specific aims: 1) Identify the transcription factor network that regulates ESX-1-dependent gene expression. 2) Define the genes required for ESX-1-dependent signal transduction. Under the first aim, the applicant proposes to use genetic, molecular and biochemical approaches to identify transcription factors and genes in the tran- scriptional network that responds to the ESX-1 translocon. Under the second aim, the applicant proposes to use genetic and molecular approaches to identify and characterize genes and signaling pathways that connect the translocon in the cytoplasmic membrane to changes in gene expression. The expected outcome following the successful completion of the proposed aims will be to define factors and pathways required for ESX-1-dependent changes in mycobacterial gene expression. This contribution will be significant because it will provide fundamen- tal insight into the regulation of bacterial factors that promote interaction with the host during the initial phases of infection, positively impacting our ability to prevent diagnose and treat TB. The proposed research is technically and conceptually innovative. Previous work on ESX-1 has focused on the mechanisms of secretion by and the effects of the ESX-1 system on the host. The proposed work focuses on the role of the ESX-1 system in regu- lating transcriptional response networks, which has not been previously considered. To the applicant’s knowledge, considering the assembly of the ESX-1 translocon as a signal that elicits changes in gene expression is a new idea. Finally, the combined multi-pronged approach to identify the mechanisms connecting the ESX-1 translocon to gene expression is innovative. Defining the molecular mechanisms underlying the control of gene expression by the ESX- translocon will advance the field by providing new mechanisms regulating mycobacterial gene expression.

项目概要：申请人的实验室发现ESX-1易位子在细胞质膜中的组装申请人的长期目标是了解其生物学意义。申请人提出ESX-1反式的组装。 Locon 引发将吞噬体裂解和细胞质暴露适应相结合的基因表达途径。该提案的总体目标是探索 ESX-1 易位子如何调节基因表达，这将有助于实现申请人的长期目标。核心假设是存在一个不寻常的特征。将 ESX-1 易位子与基因表达的广泛变化连接起来的特化转录反应网络 sion，可能将吞噬体裂解和细胞质适应联系起来。完成拟议的工作将深入了解连接吞噬体裂解的基本机制 ESX-1 的膜与基因表达的变化将通过以下这些进行检验。具体目标： 1) 确定调节 ESX-1 依赖性基因表达的转录因子网络 2)。定义 ESX-1 依赖性信号转导所需的基因在第一个目标下，申请人提出建议。使用遗传、分子和生化方法来识别转录因子和基因响应ESX-1易位子的脚本网络在第二个目标下，申请人建议使用。遗传和分子方法来识别和表征连接基因和信号通路细胞质膜中的易位子导致基因表达的变化。成功完成拟议目标将是定义 ESX-1 依赖所需的因素和途径分枝杆菌基因表达的变化这一贡献将是重要的，因为它将提供基础。深入了解细菌因子在初始阶段促进与宿主相互作用的调节感染，对我们预防诊断和治疗结核病的能力产生积极影响。拟议的研究在技术上是可行的。之前关于 ESX-1 的工作主要集中在 ESX-1 的分泌机制和概念上的创新。 ESX-1 系统对主机的影响拟议的工作重点是 ESX-1 系统在调节中的作用。转录反应网络，这是申请人之前没有考虑过的。知识，将 ESX-1 易位子的组装视为引发基因表达变化的信号最后，多管齐下的方法来识别连接ESX-1的机制。基因表达的易位子是创新的，它定义了基因控制的分子机制。 ESX-易位子的表达将通过提供调节分枝杆菌的新机制来推动该领域的发展基因表达。