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 弯曲杆菌空肠是美国及世界各地胃肠炎的主要细菌，有140个 全世界有百万例病例，每年美国有130万例Jejuni肠胃炎病例。这种感染导致 每年约30,000人死亡，主要是在欠发达国家<5岁的儿童。大多数案例。 空肠疾病是零星的，是由于污染的食物（尤其是家禽）和环境暴露而引起的 水。一些旋转的C. jejuni感染（〜1/1,000）导致Guillain-Barré综合征的发展，领先 世界上急性瘫痪的原因，以及反应性关节炎和其他后遗症。尽管患病率很高 弯曲杆菌疾病和大约40年的研究，梭状芽胞杆菌引起疾病的机制 保持不完全理解并严重理解。 细菌调节蛋白天冬氨酸残留物的磷酸化是一种众所周知的机制 介导基因表达的变化。但是，越来越多地认识到蛋白质的磷酸化 在丝氨酸/苏氨酸（S/T）上，保留也可以改变蛋白质功能。 S/T磷酸化系统使用特定 激酶以磷酸化蛋白质和磷酸酶从S/T保留中去除磷酸盐，以响应 到环境信号。重要的是，这发生在细菌病原体中，改变了许多病毒相关的 特征。我们的初步研究首次表明，提供了新的证据表明Jejuni S/T 磷酸化八种与运动/趋化性关键致病决定剂有关的蛋白质，表明 S/T磷酸化调节了空肠梭菌病毒。我们将定义C中的S/T磷酸化系统。 Jejuni，根据以下假设： 总体假设：空肠梭菌磷酸化S/T处的蛋白质保留以调节运动/趋化性和趋化性和趋化性 相关的发病机理特征，通过新型的S/T激酶/磷酸酶。 我们提出了一项详细研究的蛋白质S/t磷酸化链球菌的磷酸化，重点是该机制 运动/趋化性蛋白的磷酸化影响其功能，并确定 影响S/T磷酸化水平的蛋白质。我们将结合遗传，生化和 先进的磷蛋白学方法实现了这两个特定目的中概述的目标：目标1。 确定S/T磷酸化对运动/趋化性相关蛋白功能的功能，并AIM 2。 确定假定的S/T激酶/磷酸酶蛋白在修饰Jejuni蛋白质中的作用。在一起，这些 将为我们的长期目标奠定基础 人类中的磷酸化和中断宿主的传播和发病机理。