Establishment of an aqueous environment as a novel mechanism of bacterial pathogenesis

建立水环境作为细菌发病机制的新机制

• 批准号: 10267699
• 负责人: SHENG YANG HE
• 金额: $ 39.51万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-21 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10267699
• 关键词: 26S proteasome; ADP-Ribosylation Factors; Affect; Air; Animals; Arabidopsis; Bacteria; Bacterial Infections; Biochemical Genetics; Biochemistry; Biological; Cell membrane; Cells; Cellular biology; Communicable Diseases; Development; Dimensions; Disease; Early Endosome; Environment; Epidermis; Family; Gases; Genetic; Goals; Golgi Apparatus; Guanine Nucleotide Exchange Factors; Homeostasis; Human; Immunity; Infection; Infection Control; Innate Immune Response; Kidney; Knowledge; Laboratories; Lead; Link; Malignant Neoplasms; Mammals; Measures; Mediating; Methods; Michigan; Microbe; Modeling; Molecular Genetics; Mouse-ear Cress; Mucous body substance; Natural Immunity; Nature; Organ; Pathogenesis; Pathway interactions; Phosphorylation; Plant Leaves; Plants; Principal Investigator; Proliferating; Proteins; Pseudomonas syringae; Regulation; Research; Respiratory System; Role; Skin; Testing; Tissues; Type III Secretion System Pathway; Universities; Vesicle; Virulence; Water; Yang; aqueous; calcium-dependent protein kinase; enteropathogenic Escherichia coli; extracellular; host microbiome; host-microbe interactions; in vivo; innovation; insight; microbial; microbiome; novel; pathogen; pathogenic bacteria; prevent; ubiquitin-protein ligase; water channel

Establishment of an aqueous environment as a novel mechanism of bacterial pathogenesis PI: HE, Sheng Yang; Michigan State University Project summary Many bacterial pathogens of plants and mammals, including humans, use the highly conserved type III secretion system (T3SS) to inject “effector proteins” into the host cell as an important paradigm of pathogenesis. The long-term goal of this project is to characterize a newly discovered T3SS-mediated virulence strategy by which bacterial pathogens create an aqueous extracellular environment in host tissues. In nature, many host-pathogen and host-microbiome interactions occur in air-exposed/connected host organs/tissues (e.g., epidermis/skin and gas-exchange organs including respiratory systems and plant leaves) in which water availability is limited and/or variable. Because microbes generally require a moist/aqueous/mucous environment to survive and proliferate, it is not well understood whether microbes actively establish an infection-conducive aqueous environment in host organs. In humans, malfunction of aquaporins has been associated with infectious diseases, kidney malfunction and even cancer development and there is an emerging link between aquaporin- mediated water transport and pathogenesis of enteropathogenic Escherichia coli. However, cause-effect relationships often remain unclear. In the past 25 years, the Principal Investigator’s lab has used the model Arabidopsis thaliana – Pseudomonas syringae interaction to discover and characterize T3SS-mediated bacterial infection mechanisms. By taking advantage of the genetic tractability of Arabidopsis and a well-characterized T3SS effector repertoire in P. syringae, the PI’s lab recently discovered a critical role of an aqueous environment in bacterial pathogenesis. In this application, three specific aims are proposed to test the central hypothesis that, by altering (i) ARF-GEFMIN7-dependent vesicular traffic and (ii) phosphorylation of aquaporins involved in regulating water transport across host plasma membrane, P. syringae disrupts water homeostasis across the host plasma membrane, resulting in an aqueous extracellular environment as an important mechanism of pathogenesis. Aim 1 will determine the role of ARF-GEFMIN7-associated host proteins in regulating vesicular traffic of aquaporins and extracellular water. Aim 2 will investigate how P. syringae T3SS effector proteins target ARF-GEFMIN7- associated vesicle traffic and aquaporins to induce an aqueous extracellular environment. Aim 3 will elucidate how activation of host immunity prevents the virulence actions of P. syringae T3SS effectors as a novel dimension of the host innate immune response. Contemporary methods in molecular genetics, cell biology, biochemistry and microbial pathogenesis will be used in this study. Successful completion of this research will significantly advance our understanding of a newly discovered bacterial virulence mechanism and its interplay with host innate immunity. As many host-microbe interactions occurs in air-connected host organs/tissues, in which water availability is restricted, it is hoped that this original research will stimulate studies to broadly examine water regulation in other host-pathogen interactions, and, in the long- term, facilitate the development of innovative and broadly applicable measures for controlling infectious diseases in diverse eukaryotic hosts, including plants, animals and humans.

水环境作为细菌致病新机制的建立 PI：HE，Sheng Yang;密歇根州立大学 项目摘要 植物和哺乳动物包括人类的许多细菌病原体使用高度保守的III型分泌系统 （T3 SS）将“效应蛋白”注射到宿主细胞中作为发病机制的重要范例。长期目标是 该项目是表征一种新发现的T3 SS介导的毒力策略，通过该策略，细菌病原体产生一种 宿主组织中的含水细胞外环境。在自然界中，许多宿主-病原体和宿主-微生物组相互作用发生 在空气暴露/连接的宿主器官/组织中（例如，表皮/皮肤和气体交换器官，包括呼吸系统 和植物叶片）中的水分可利用性是有限的和/或可变的。因为微生物通常需要 潮湿/水性/粘液环境中生存和增殖，微生物是否积极建立 在宿主器官中有利于感染的水环境。在人类中，水通道蛋白的功能障碍与 感染性疾病，肾功能障碍，甚至癌症的发展，水通道蛋白- 介导的水转运和致病性肠致病性大肠杆菌。然而，因果关系往往 仍然不清楚。在过去的25年里，首席研究员的实验室使用了拟南芥-假单胞菌的模型 发现和表征T3 SS介导的细菌感染机制。通过利用 拟南芥的遗传易处理性和一个充分表征的T3 SS效应器库在P. lingae，PI的实验室最近 发现了水环境在细菌发病机制中的关键作用。在本申请中，三个具体目标是 提出测试中心假设，即通过改变（i）ARF-GEFMIN 7依赖的囊泡运输和（ii）磷酸化 在参与调节水跨宿主质膜运输的水通道蛋白中，P. lingae破坏了水的稳态 穿过宿主质膜，导致水性细胞外环境作为一种重要的机制， 发病机制目的1将确定ARF-GEFMIN 7相关宿主蛋白在调节细胞囊泡运输中的作用。 水通道蛋白和细胞外水。目的2将研究P. lingae T3 SS效应蛋白如何靶向ARF-GEFMIN 7 - 1。 相关的囊泡运输和水通道蛋白，以诱导水性细胞外环境。目标3将阐明如何 宿主免疫的激活阻止了作为宿主的新维度的P. erichingae T3 SS效应子的毒力作用 先天免疫反应分子遗传学、细胞生物学、生物化学和微生物 发病机制将用于本研究。这项研究的成功完成将大大促进我们的理解 新发现的细菌毒力机制及其与宿主先天免疫的相互作用。宿主微生物 相互作用发生在与空气相连的宿主器官/组织中，其中水的可用性受到限制，希望这种原始的相互作用可以在体内进行。 研究将刺激研究，以广泛研究水调节在其他宿主-病原体相互作用，并在长期- 促进制定创新和广泛适用的措施，以控制不同国家的传染病。 真核宿主，包括植物、动物和人类。