Understanding the role of Ngo1049 in subverting host-mediated metal starvation in Neisseria gonorrhoeae

了解 Ngo1049 在颠覆淋病奈瑟菌宿主介导的金属饥饿中的作用

• 批准号: 10385981
• 负责人: Ian Kimani Liyayi
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10385981
• 关键词: Affect; Antibiotic Resistance; Bacterial Antibiotic Resistance; Bacterial Physiology; Binding; Biochemistry; Bioinformatics; Cells; Collaborations; Data; Development; Epithelial; Epithelial Cells; Exposure to; Genes; Gonorrhea; Growth; Human; Immune; Immunity; Infection; Iron; Laboratory Research; Lactoferrin; Leukocyte L1 Antigen Complex; Manganese; Mediating; Membrane Transport Proteins; Metals; Microbiology; Mucous Membrane; Neisseria; Neisseria gonorrhoeae; Nutrient; Nutritional Immunity; Pathogenesis; Pathogenicity; Proteins; Public Health; Regulation; Regulon; Role; Sexually Transmitted Diseases; Site; Starvation; Surface; System; Testing; Transcript; Transferase; Transferrin; Translating; Vaccine Therapy; Vaccines; Work; Zinc; bacterial genetics; base; career; combat; drug resistant bacteria; extracellular; gene product; human pathogen; in vivo; macrophage; member; metal chelator; neutrophil; new therapeutic target; novel; novel therapeutics; pathogen; pathogenic bacteria; periplasm; psoriasin; training project; uptake; zinc-binding protein

Abstract Neisseria gonorrhoeae (Ngo) is an obligate human bacterial pathogen that causes the sexually transmitted infection gonorrhea. Due to increasing rates of gonorrhea and increased antibiotic resistance, vaccines and new therapeutics are urgently needed. A promising strategy is targeting nutrient metal acquisition systems since they are usually conserved, expressed during infection, and essential for bacterial survival in vivo. Ngo undermines host metal restriction mechanisms by expressing outer membrane transporters to acquire essential metals from human metal-sequestering proteins such zinc from calprotectin and psoriasin and iron from lactoferrin and transferrin. However, many gene products that support Ngo growth in metal-limiting conditions remain uncharacterized. We found that the product of the ngo1049 gene is highly expressed in Ngo grown under zinc- limiting conditions. Ngo1049 is conserved among pathogenic Neisseria, and bioinformatic analysis predicts that Ngo1049 is a metal-binding transferase localized in the periplasm. A Zur binding motif was identified upstream of ngo1049, suggesting expression is regulated by Zur (zinc uptake regulator), which represses expression in high zinc concentrations. ngo1049 transcripts are highly induced during Ngo infection of human endocervical cells, indicating a potential role for Ngo1049 in Ngo pathogenesis. Based on these findings, I hypothesize Ngo1049 is a Zur-regulated protein that facilitates zinc acquisition in metal-limited conditions at inflamed epithelial surfaces. To test this hypothesis, in this F31-Diversity submission I propose to determine the localization and regulation mechanism(s) of Ngo1049. Second, I will examine the contribution of Ngo1049 to zinc acquisition in metal-limiting conditions. Lastly, I will define how Ngo1049 enables Ngo survival at inflamed mucosal surfaces after exposure to epithelial cells and human immune cells that contain metal chelating proteins. By defining this new member of the Ngo metal regulon, my work will potentially point to new therapies for this antibiotic-resistant bacterium. Through its combination of bacterial physiology and genetics, biochemistry, and cellular microbiology, as well as the professional development opportunities available to me during my graduate training, this project will provide me with the background and expertise to pursue a career as the leader of an academic research laboratory in host-pathogen interactions.

抽象的 淋病奈瑟菌 (Ngo) 是一种专性人类细菌病原体，可引起性传播疾病 感染淋病。由于淋病发病率上升和抗生素耐药性增加，疫苗和新药物 迫切需要治疗。一个有前景的策略是针对营养金属获取系统，因为它们 通常是保守的，在感染过程中表达，并且对于细菌在体内的生存至关重要。非政府组织破坏 通过表达外膜转运蛋白来获取必需金属的宿主金属限制机制 人体金属螯合蛋白，例如钙卫蛋白和牛皮癣素中的锌以及乳铁蛋白中的铁和 转铁蛋白。然而，许多支持 Ngo 在金属限制条件下生长的基因产物仍然存在 没有特征的。我们发现ngo1049基因的产物在锌环境下生长的Ngo中高度表达 限制条件。 Ngo1049 在致病性奈瑟菌中是保守的，生物信息学分析预测 Ngo1049 是一种位于周质的金属结合转移酶。在上游鉴定出 Zur 结合基序 ngo1049 的表达，表明表达受到 Zur（锌摄取调节剂）的调节，Zur 抑制表达 高锌浓度。 ngo1049 转录本在 Ngo 感染人类宫颈管过程中被高度诱导 细胞，表明 Ngo1049 在 Ngo 发病机制中的潜在作用。根据这些发现，我假设 Ngo1049 是一种 Zur 调节蛋白，可促进炎症时金属限制条件下锌的获取 上皮表面。为了检验这个假设，在这份 F31-Diversity 提交中，我建议确定 Ngo1049 的本地化和调控机制。其次，我将考察Ngo1049对锌的贡献 在金属限制条件下进行采集。最后，我将定义 Ngo1049 如何使 Ngo 在发炎的情况下生存 暴露于含有金属螯合蛋白的上皮细胞和人体免疫细胞后的粘膜表面。 通过定义 Ngo 金属调节子的这个新成员，我的工作可能会为此找到新的疗法 抗生素耐药细菌。通过细菌生理学和遗传学、生物化学和 细胞微生物学，以及我在研究生期间可获得的专业发展机会 培训，该项目将为我提供背景和专业知识，以追求作为领导者的职业生涯 宿主与病原体相互作用的学术研究实验室。