Mechanistic evaluation of resistance to sulfite toxicity in Salmonella

沙门氏菌抗亚硫酸盐毒性的机制评价

• 批准号: 10724560
• 负责人: Johanna Rebecca Elfenbein
• 金额: $ 24.75万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-06 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10724560
• 关键词: Amino Acids; Anabolism; Anti-Bacterial Agents; Area; Bacteria; Bacterial Infections; Binding; Biogenesis; Cause of Death; Cell physiology; Cessation of life; ChIP-seq; Cysteine; DNA; Data; Disease; Drug Metabolic Detoxication; Enteral; Evaluation; Flagella; Food; Future; Gastroenteritis; Gene Expression; Genes; Genetic Transcription; Growth; Homologous Gene; Immune; Immune response; In Vitro; Infection; Inflammation; Innate Immune Response; Innate Immune System; Intestines; Libraries; Life; Link; Lipids; Lipopolysaccharides; Liver; Mediating; Methionine; Microbe; Modeling; Mus; Neutrophil Infiltration; Organ; Orthologous Gene; Outcome; Pathogenesis; Phase; Play; Process; Production; Proteins; Pseudomonas aeruginosa; Publishing; Reaction; Regulation; Regulon; Repression; Resistance; Role; Salmonella; Salmonella enterica; Salmonella infections; Sepsis; Side; Signaling Molecule; Spleen; Stimulus; Stress; Sulfate; Sulfite reductase; Sulfites; Sulfur; Systemic infection; Testing; Toxic effect; United States; Vibrio cholerae; Virulence; Virulence Factors; Work; Yersinia enterocolitica; amino acid metabolism; antimicrobial; biological adaptation to stress; combat; density; enteric infection; enteric pathogen; foodborne; foodborne illness; foodborne pathogen; gene repression; genetic approach; gut colonization; high reward; high risk; human pathogen; in vivo; mouse model; mutant; neutrophil; non-typhoidal Salmonella; novel therapeutic intervention; pathogen; pathogenic bacteria; prevent; promoter; response; transcriptome

PROJECT SUMMARY Sulfites are reactive sulfur species with ubiquitous distribution in all life forms. They are byproducts of sulfated amino acid metabolism and have been used for decades to limit bacterial growth in food products. Because sulfites can react with proteins, DNA, and lipids to disrupt cellular processes, detoxification of sulfites is essential for cellular viability. Interestingly, sulfites are produced by activated neutrophils in response to bacterial infection, indicating they may play an important role in host-pathogen interactions. The purpose of this work is to establish how sulfites impact the outcome of bacterial infection, using Salmonella enterica as a model pathogen. Salmonella enterica is a leading cause of bacterial foodborne gastroenteritis and is the leading cause of death from foodborne disease in the United States. In our published work, we characterized the role of a transcriptional regulator, YeiE, in Salmonella pathogenesis. We demonstrated that yeiE is critical for Salmonella to colonize the intestine because it regulates production of flagella, which are a key virulence factor required for enteric salmonellosis. Recent work in Cronobacter sakazakii demonstrated that YeiE senses excess sulfite to regulate expression of a sulfite reductase and is needed to survive neutrophil killing. YeiE homologs are present in many bacterial pathogens, suggesting an important role for sulfite sensing by YeiE in host-pathogen interactions. We hypothesize that YeiE regulates the sulfite stress response to allow Salmonella to withstand innate immune control. In Aim 1, we will determine the role of sulfite reduction in salmonellosis. We will establish the impact of YeiE-regulated sulfite reduction in murine infection models of gastroenteritis and sepsis. Since activated neutrophils produce sulfite and neutrophilic inflammation is a key to host control of Salmonella, it is important to understand how YeiE mediates survival in the face of host-derived sulfite stress. In Aim 2, we will establish the mechanism by which YeiE regulates resistance to sulfite toxicity. We will establish the genes regulated by sulfite-bound YeiE using chromatin immunoprecipitation-sequencing and will establish the genes needed to withstand sulfite stress using a high-density transposon mutant library. Combined analysis of these two unbiased approaches will allow us to establish how YeiE coordinates the bacterial response to sulfite stress. YeiE homologs are distributed amongst many diverse bacterial pathogens, so our work will have application to a broad range of host-pathogen interactions. Successful completion of this work will demonstrate how bacteria resist sulfite stress in vivo and will lead to future work to investigate the role of sulfite as a cross- kingdom signaling molecule and its impact on the immune response to bacterial infections.

项目摘要 亚硫酸盐是一种广泛存在于所有生命形式中的活性硫物种。它们是硫酸化的副产品 氨基酸代谢，并且几十年来一直用于限制食品中的细菌生长。因为 亚硫酸盐可以与蛋白质，DNA和脂质反应，破坏细胞过程，亚硫酸盐的解毒至关重要 细胞存活率。有趣的是，亚硫酸盐是由活化的嗜中性粒细胞在细菌感染后产生的， 表明它们可能在宿主-病原体相互作用中起重要作用。这项工作的目的是 确定亚硫酸盐如何影响细菌感染的结果，使用沙门氏菌作为模型 病原体肠道沙门氏菌是细菌性食源性胃肠炎的主要原因， 在美国死于食源性疾病的人数。在我们发表的工作中，我们描述了一个 转录调节因子YeiE在沙门氏菌致病机制中的作用。我们证明了yeiE对沙门氏菌至关重要 因为它调节鞭毛的产生，这是一个关键的毒力因子， 肠道沙门氏菌病最近在坂崎县的研究表明，YeiE感觉到过量的亚硫酸盐， 调节亚硫酸盐还原酶的表达，并且是嗜中性粒细胞杀伤存活所需的。存在YeiE同源物 在许多细菌病原体中，表明YeiE对亚硫酸盐的敏感性在宿主-病原体中起重要作用 交互.我们假设YeiE调节亚硫酸盐应激反应，使沙门氏菌能够 抵抗先天免疫控制在目标1中，我们将确定亚硫酸盐还原在沙门氏菌病中的作用。我们 将确定YeiE调节的亚硫酸盐还原在胃肠炎的鼠感染模型中的影响， 败血症由于活化的中性粒细胞产生亚硫酸盐和嗜酸性炎症是宿主控制炎症的关键。 沙门氏菌，重要的是要了解如何YeiE介导的生存在面对宿主来源的亚硫酸盐应激。在 目的2：研究叶叶绿素对亚硫酸盐胁迫抗性的调控机制.我们将建立 亚硫酸盐结合的YeiE调控的基因，使用染色质免疫沉淀测序，并将建立 使用高密度转座子突变体库来抵抗亚硫酸盐胁迫所需的基因。组合分析 这两种无偏倚的方法将使我们能够建立YeiE如何协调细菌对 亚硫酸盐胁迫YeiE同源物分布在许多不同的细菌病原体中，因此我们的工作将具有 应用于广泛的宿主-病原体相互作用。这项工作的成功完成将证明 细菌如何抵抗体内亚硫酸盐的压力，并将导致未来的工作，以调查亚硫酸盐的作用，作为一个交叉- 王国信号分子及其对细菌感染免疫反应的影响。