Protein phosphorylation and Campylobacter jejuni pathogenesis

蛋白质磷酸化和空肠弯曲菌发病机制

基本信息

批准号：
10448142
负责人：
STUART A THOMPSON
金额：
$ 23.1万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-11 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10448142
关键词：
5 year old Acute Acute Diarrhea Adherence Affect Anabolism Aspartate Bacteria Bacterial Physiology Biochemical Biological Assay Birds Campylobacter Campylobacter jejuni Cattle Cell Wall Cells Cessation of life Characteristics Chemoreceptors Chemotaxis Child Complex Country Data Defect Development Disease Domestic Fowls Environment 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 Serine/Threonine Phosphatase Protein phosphatase Protein-Serine-Threonine Kinases Proteins Refrigeration Reiter Disease Research Role Rotation Route Serine Serine/Threonine Phosphorylation Signal Transduction Site-Directed Mutagenesis Swimming System Tertiary Protein Structure Testing Threonine Threonine Phosphorylation Site Time Transducers Virulence Virulence Factors Water Work bacterial metabolism base cell motility druggable target extracellular genetic regulatory protein host colonization in vivo novel pathogen pathogenic bacteria phosphoproteomics prevent protein function response transmission process wild bird

项目摘要

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。每年有超过 30,000 人死亡，主要发生在不发达国家的 5 岁以下儿童中。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。尽管患病率很高，但它是世界上急性瘫痪以及反应性关节炎和其他后遗症的原因。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) 残基也改变蛋白质功能。S/T 磷酸化系统使用特定的功能。作为响应，激酶磷酸化蛋白质，磷酸酶去除 S/T 残基中的磷酸盐重要的是，这种情况发生在细菌病原体中，改变了许多与毒力相关的因素。我们的初步研究首次表明空肠弯曲菌 S/T 提供了新的证据。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定义空肠弯曲菌中的 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/趋化性和相关的发病机制特征，通过新型 S/T 激酶/磷酸酶。我们建议对空肠弯曲菌中的蛋白质 S/T 磷酸化进行详细研究，重点关注其机制运动性/趋化性相关蛋白的磷酸化影响其功能，并决定我们将结合遗传、生物化学和影响 S/T 磷酸化水平的蛋白质。先进的磷酸化蛋白质组学方法可实现以下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。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。