Vibrio vulnificus toxin-receptor interactions

创伤弧菌毒素受体相互作用

基本信息

批准号：
10198737
负责人：
Karla J F Satchell
金额：
$ 19.94万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-19 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10198737
关键词：
Address Bacteria Binding Blood Circulation C-terminal CRISPR screen Calcium Caspase Cell membrane Cell surface Cells Cessation of life Chromosomes Climate Clinical Consumption Cytoskeleton Cytosol Cytotoxin Death Rate Disease Eating Economic Burden Epithelial Eukaryotic Cell Family Fishes Food Food Contamination Functional disorder Future Gene Expression Regulation Genes Genetic Screening Glycolipids Golgi Apparatus Grant Growth Harvest Human Incidence Infection Inflammatory Insecta Integration Host Factors Intestines Intoxication Ions Knowledge Lead Lethal Dose 50 Life Mexico Morbidity - disease rate Mus Mutation N-terminal Nature Operon Patients Phytic Acid Policies Polysaccharides Protease Domain Proteins Receptor Cell Resistance Route Seafood Seawater Sepsis Septicemia Shellfish Structure Surface Swimming Terminal Repeat Sequences Testing Toxin Variant Vibrio Vibrio cholerae Vibrio vulnificus Virulence Virulence Factors alpha Toxin arm base beta-Lactamase cytokine cytotoxic foodborne pathogen genetic selection human pathogen inhibition of autophagy intestinal epithelium mortality mouse model mutant pathogenic bacteria receptor receptor function transmission process

项目摘要

ABSTRACT Vibrio vulnificus is a Gram-negative bacterial pathogen that causes severe life-threatening infections in humans after eating shellfish or swimming in warm seawater. In the US, there are about 200 cases annually with high rates of mortality and morbidity. While the number of cases is low compared to other food-borne pathogens, the high rates of death among patients exerts a significant burden on human life and the economy. This is expected to increase as incidence of V. vulnificus infections is rising due to the climate crisis. The most prominent virulence factor of V. vulnificus is a 5208 aa MARTX family toxin that delivers cytotoxic effectors to cells and these effectors are essential for virulence in a mouse model. While extensive information is known about the mechanisms of the effectors, very little is known about the remaining over 2800 aa of the protein toxin. These regions are known to be both necessary and sufficient for secretion of the toxin from the bacteria, interaction of the toxin with the eukaryotic cell surface, and translocation of the effectors across the plasma membrane. The project proposes to identify host cell receptors and host factors essential for intoxication using a genetic screen and selection for mutant cells that survive the cytolytic action of the toxin. We will also use a structure-function based approach to identify regions of the MARTX toxin essential for Type I secretion from the bacterium, binding of the toxin to a putative surface receptor, formation of a pore in the plasma membrane, and translocation of effectors to the eukaryotic cell cytosol. This study will advance our understanding of the function of the MARTX toxin repeat regions in detail and is expected to impact our understanding of pathophysiology during V. vulnificus infection.

抽象的创伤弧菌是一种革兰氏阴性细菌病原体，可导致严重危及生命的感染人类食用贝类或在温暖的海水中游泳后。在美国，每年约有200起病例死亡率和发病率很高。虽然与其他食源性疾病相比，病例数较低由于病原体的存在，患者的高死亡率给人类生命和经济带来了沉重负担。由于气候危机，创伤弧菌感染的发病率不断上升，预计这一数字还会增加。最创伤弧菌的重要毒力因子是一种 5208 个氨基酸的 MARTX 家族毒素，可将细胞毒性效应子传递给细胞和这些效应子对于小鼠模型的毒力至关重要。虽然已知大量信息关于效应子的机制，人们对蛋白质剩余的 2800 多个氨基酸知之甚少毒素。已知这些区域对于细菌分泌毒素是必要且充分的，毒素与真核细胞表面的相互作用，以及效应器跨血浆的易位膜。该项目提议利用对在毒素的细胞溶解作用中存活下来的突变细胞进行遗传筛选和选择。我们还将使用一个基于结构功能的方法来识别 I 型分泌所必需的 MARTX 毒素区域细菌，毒素与假定的表面受体结合，质膜上形成孔，以及效应子易位至真核细胞胞质。这项研究将加深我们对详细介绍了 MARTX 毒素重复区域的功能，预计将影响我们对创伤弧菌感染期间的病理生理学。