Protein phosphorylation and Campylobacter jejuni pathogenesis

蛋白质磷酸化和空肠弯曲菌发病机制

• 批准号: 10608212
• 负责人: STUART A THOMPSON
• 金额: $ 19.25万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-11 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10608212
• 关键词: 5 year old; Acute; Acute Diarrhea; Adherence; Affect; Anabolism; Aspartate; Bacteria; Bacterial Physiology; Binding; Biochemical; Biological Assay; Birds; Campylobacter; Campylobacter jejuni; Cattle; Cell Wall; Cells; Cessation of life; Characteristics; Chemoreceptors; Chemotaxis; Child; Communication; Complex; Cytoplasm; Data; Defect; Developing Countries; Development; Disease; Domestic Fowls; Environment; Environmental Pollution; Exposure to; Flagella; Food Contamination; Food Processing; Freezing; Gastroenteritis; Gene Expression; Gene Expression Regulation; Genetic; Goals; Guillain Barré Syndrome; High Prevalence; Human; Infection; Ingestion; Interruption; Intervention; Intestines; Invaded; Lead; Maps; Meat; Mediating; Membrane; Metabolism; Microbial Biofilms; Movement; Mucous body substance; Mutation; Mutation Analysis; Nature; Organelles; Paralysed; Pathogenesis; Pathogenicity; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphoserine; Phosphotransferases; Process; Production; Protein Dephosphorylation; Protein phosphatase; Protein-Serine-Threonine Kinases; Proteins; Recreation; Refrigeration; Reiter Disease; Research; Role; Rotation; Route; Serine; Serine/Threonine Phosphorylation; Signal Transduction; Site-Directed Mutagenesis; Swimming; System; Testing; Threonine; Threonine Phosphorylation Site; Time; Transducers; Virulence; Virulence Factors; Viscosity; Water; Work; bacterial metabolism; cell motility; druggable target; extracellular; genetic regulatory protein; host colonization; in vivo; migration; novel; pathogen; pathogenic bacteria; phosphoproteomics; prevent; protein function; response; transmission process; virulence gene; wild bird

ABSTRACT Campylobacter jejuni is a primary bacterial cause of gastroenteritis in the US and throughout the world, with 140 million cases worldwide and 1.3 million cases of C. jejuni gastroenteritis in the US each year. This infection leads to >30,000 deaths annually, primarily in children <5 years old in underdeveloped countries. Most cases of C. jejuni disease are sporadic and result from contaminated food (especially poultry) and exposure to environmental waters. Some C. jejuni infections (~1/1,000) lead to the development of Guillain-Barré Syndrome, the leading cause of acute paralysis in the world, as well as reactive arthritis and other sequelae. Despite the high prevalence of Campylobacter disease and some 40 years of research, the mechanisms by which C. jejuni causes disease remain incompletely understood and severely understudied. Phosphorylation of aspartate residues of bacterial regulatory proteins is a well-known mechanism for mediating changes in gene expression. However, it is increasingly recognized that phosphorylation of proteins at serine/threonine (S/T) residues also modifies protein function. The S/T phosphorylation system uses specific kinases to phosphorylate proteins, and phosphatases to remove the phosphates from S/T residues, in response to environmental signals. Importantly, this occurs in bacterial pathogens, modifying many virulence-associated characteristics. For the first time, our preliminary studies indicate provide new evidence that C. jejuni S/T phosphorylates eight proteins related to the critical pathogenic determinants of motility/chemotaxis, suggesting that S/T phosphorylation modulates C. jejuni virulence. We will define the S/T phosphorylation system in C. jejuni, according to the following hypothesis: Overall Hypothesis: C. jejuni phosphorylates proteins at S/T residues to modulate motility/chemotaxis and related pathogenesis characteristics, by means of novel S/T kinases/phosphatases. We propose a detailed study of protein S/T phosphorylation in C. jejuni, focusing on the mechanism by which the phosphorylation of motility/chemotaxis-related proteins affects their functions, as well as determining the proteins that affect the levels of S/T phosphorylation. We will use a combination of genetic, biochemical, and advanced phosphoproteomics approaches to achieve the goals outlined in these two specific aims: Aim 1. Determine the role of S/T phosphorylation for the function of motility/chemotaxis-related proteins, and Aim 2. Determine the roles of putative S/T kinase/phosphatase proteins in modifying C. jejuni proteins. Together, these will lay the groundwork for our long-term goal of identifying druggable targets to interrupt C. jejuni S/T phosphorylation and interrupting inter-host transmission and pathogenesis in humans.

抽象的 空肠弯曲菌是美国和全世界胃肠炎的主要原因，有 140 全球每年有 100 万例空肠弯曲菌胃肠炎病例，美国每年有 130 万例空肠弯曲菌胃肠炎病例。这种感染导致 每年有超过 30,000 人死亡，主要是欠发达国家 5 岁以下的儿童。大多数情况C. 空肠病是散发性的，由受污染的食物（尤其是家禽）和暴露于环境中引起 水域。一些空肠弯曲菌感染（~1/1,000）会导致格林-巴利综合征的发生，这是主要的疾病 导致世界范围内的急性瘫痪以及反应性关节炎等后遗症。尽管患病率很高 弯曲杆菌疾病的研究和大约 40 年的研究，空肠弯曲杆菌引起疾病的机制 仍然不完全被理解并且未被充分研究。 细菌调节蛋白天冬氨酸残基的磷酸化是众所周知的机制 介导基因表达的变化。然而，人们越来越认识到蛋白质的磷酸化 丝氨酸/苏氨酸 (S/T) 残基也会改变蛋白质功能。 S/T磷酸化系统使用特定的 作为响应，激酶磷酸化蛋白质，磷酸酶去除 S/T 残基中的磷酸盐 环境信号。重要的是，这发生在细菌病原体中，改变了许多与毒力相关的 特征。我们的初步研究首次表明，提供了空肠弯曲菌 S/T 的新证据 磷酸化与运动/趋化性的关键致病决定因素相关的八种蛋白质，表明 S/T 磷酸化调节空肠弯曲菌毒力。我们将在 C 中定义 S/T 磷酸化系统。 空肠病，根据以下假设： 总体假设：空肠弯曲菌在 S/T 残基处磷酸化蛋白质以调节运动性/趋化性和 相关的发病机制特征，通过新型 S/T 激酶/磷酸酶。 我们提出了对空肠弯曲菌中蛋白质 S/T 磷酸化的详细研究，重点关注其机制 运动/趋化相关蛋白的磷酸化影响其功能，并决定 影响 S/T 磷酸化水平的蛋白质。我们将结合遗传、生化和 先进的磷酸蛋白质组学方法来实现这两个具体目标中概述的目标：目标 1。 确定 S/T 磷酸化对运动/趋化相关蛋白功能的作用，目标 2。 确定假定的 S/T 激酶/磷酸酶蛋白在修饰空肠弯曲菌蛋白中的作用。在一起，这些 将为我们确定可药物靶标以阻断空肠弯曲菌 S/T 的长期目标奠定基础 磷酸化并中断人类宿主间传播和发病机制。