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: 10293988
• 负责人: SHENG YANG HE
• 金额: $ 39.43万
• 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/10293988
• 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; 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.

建立水性环境作为细菌发病机理的新机制 PI：他，Sheng Yang；密歇根州立大学 项目摘要 包括人类在内的许多植物和哺乳动物的细菌病原体使用高度保守的III型分泌系统 （T3SS）将“效应蛋白”注入宿主细胞中，这是发病机理的重要范式。这个长期目标 项目是为了表征新发现的T3SS介导的病毒策略，细菌病原体创建一个 宿主组织中的细胞外环境。在自然界中，发生了许多宿主 - 生病和宿主 - 微生物组相互作用 在暴露/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）其中的水有限和/或可变。因为微生物通常需要 潮湿/水性/粘液环境可生存和繁殖，尚不清楚微生物是否积极建立 宿主器官中的感染传统水环境。在人类中，水通道蛋白的故障与 传染病，肾功能故障甚至癌症的发展，Aquaporin-之间存在着新的联系 介导的水运输和肠病大肠杆菌的发病机理。但是，原因效应的关系经常 保持不清楚。在过去的25年中，主要研究者的实验室使用了拟南芥模型 - 假单胞菌 丁香的相互作用以发现和表征T3SS介导的细菌感染机制。通过利用 拟南芥和特征良好的T3SS效应子曲目的遗传障碍性在P. syringae中，PI的实验室最近 发现了水性环境在细菌发病机理中的关键作用。在此应用中，三个具体目标是 提议检验中心假设，即通过改变（i）ARF-GEFMIN7依赖性囊泡流量和（ii）磷酸化。 涉及跨寄主质膜的水通道的水通道蛋白，丁香假单胞菌破坏了水稳态 穿过宿主质膜，导致水外环境作为重要机制 发病。 AIM 1将确定ARF-GEFMIN7相关宿主蛋白在控制囊泡流量中的作用 水通道和细胞外水。 AIM 2将研究丁香假单胞菌T3SS效应蛋白如何靶向ARF-GEFMIN7- 相关的囊泡交通和水通道蛋白可诱导水外环境。 AIM 3将阐明如何 宿主免疫的激活阻止丁香假单胞菌T3SS的病毒作用作为宿主的新维度 先天免疫反应。分子遗传学，细胞生物学，生物化学和微生物的当代方法 发病机理将在这项研究中使用。成功完成这项研究将大大提高我们的理解 新发现的细菌病毒机制及其与宿主先天免疫的相互作用。就像许多主机菌一样 相互作用发生在空气连接的宿主器官/组织中，在其中限制了水，希望这种原始 研究将刺激研究以广泛检查其他宿主病原体相互作用中的水调节，并在长期以来 术语，促进创新且广泛适用的措施，以控制潜水员的传染病 真核生物宿主，包括植物，动物和人类。