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

项目总结： 申请人的实验室发现，ESX-1易位子在细胞膜上的组装 调节基因表达。申请者的长期目标是了解其生物学意义 ESX-1依赖的基因表达变化。申请人提议ESX-1反式-1的组装- Locon诱导的基因表达途径结合了吞噬小体的裂解和对细胞质暴露的适应。 这项提案的总体目标是探索ESX-1易位子如何调节基因表达，这 将有助于解决申请者的长期目标。中心假设是有一种不讨人喜欢的- 将ESX-1易位子与基因表达的广泛变化连接起来的转录反应网络- Sion，潜在地将吞噬小体裂解和细胞质适应联系起来。理由是，成功的COM- 这项拟议工作的完成将使我们深入了解吞噬小体裂解的基本机制。 膜上受ESX-1基因表达的改变。核心假说将通过以下方式进行检验 具体目的：1)确定调控ESX-1依赖基因表达的转录因子网络。2) 定义依赖ESX-1的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鉴定转基因植物中的转录因子和基因。 响应ESX-1易位子的脚本网络。在第二个目标下，申请人提议使用 用遗传学和分子方法来识别和表征连接基因和信号通路的 转位蛋白在细胞质膜上对基因表达的变化。之后的预期结果是 成功完成提议的目标将确定依赖ESX-1所需的因素和途径 分枝杆菌基因表达的变化。这一贡献将是重大的，因为它将为以下方面提供基础- TAL洞察细菌因子的调节，促进与宿主的相互作用在最初的阶段 感染，对我们预防、诊断和治疗结核病的能力产生了积极影响。拟议的研究在技术上是 并在概念上有所创新。以前关于ESX-1的工作主要集中在和的分泌机制上。 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 转录反应网络的延迟，这是以前没有考虑过的。至申请人的 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，确定连接ESX-1的机制的组合多管齐下的方法 转位到基因表达是创新的。确定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这一领域的发展 基因表达。

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.