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

• 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
• 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

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 调节基因表达。申请人的长期目标是了解 基因表达中的ESX-1依赖性变化。申请人建议ESX-1反式- locon eligible基因表达途径，将吞噬体溶解和适应与细胞质暴露相结合。 本研究的总体目标是探索ESX-1易位子如何调节基因表达， 有助于实现申请人的长期目标。核心假设是有一个uncharac- 将ESX-1转位子与基因表达的广泛变化联系起来的标准化转录反应网络， 锡永将吞噬体溶解和细胞质适应联系起来。理由是，成功的COM- 所提出的工作的一部分将提供深入了解的基本机制，连接裂解的吞噬体 ESX-1对细胞膜的影响及基因表达的变化。中心假设将通过以下方式进行检验： 具体目标：1）鉴定调节ESX-1依赖性基因表达的转录因子网络。（二） 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 利用遗传学、分子生物学和生物化学方法来鉴定转录因子和基因， 响应ESX-1 translocon的脚本网络。根据第二个目标，申请人建议使用 基因和分子方法来识别和表征基因和信号通路， 细胞质膜中的易位子与基因表达的变化有关。预期成果 成功完成拟议目标的关键是确定ESX-1依赖型 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- 深入了解细菌因子的调节，促进与宿主在初始阶段的相互作用， 感染，积极影响我们预防诊断和治疗结核病的能力。这项研究在技术上 在概念上创新。以前对ESX-1的研究主要集中在细胞分泌的机制上， ESX-1系统对主机的影响。建议的工作重点是ESX-1系统在regu中的作用， 转录反应网络，这是以前没有考虑过的。申请人的 知识，认为ESX-1易位子的组装是引起基因表达变化的信号 是一个新的想法。最后，结合多管齐下的方法来确定连接ESX-1的机制 转座基因表达是创新的。定义基因调控的分子机制 通过ESX-易位子的表达将通过提供调节分枝杆菌的新机制来推进该领域 基因表达。