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 Neisseria Gonorrhoeae（NGO）是一种强制性的人类细菌病原体，导致性传播 感染淋病。由于淋病的速率增加和抗生素耐药性，疫苗和新的 迫切需要治疗。有希望的策略是针对营养金属采集系统 通常是保守的，在感染期间表达，对于体内细菌生存至关重要。非政府组织破坏了 宿主金属限制机制通过表达外膜转运蛋白从 人金属序列蛋白来自钙染色蛋白和牛皮癣的锌和乳铁蛋白的铁和铁和铁 转铁蛋白。但是，许多支持金属限制条件下非政府组织增长的基因产品仍然存在 未表征。我们发现，NGO1049基因的乘积在锌下种植的非政府组织高度表达 限制条件。 NGO1049在致病性奈瑟菌中保守，生物信息学分析预测， NGO1049是局部局部局部的金属结合转移酶。在上游确定了Zur结合基序 NGO1049的表达是由Zur（锌摄取调节剂）调节的，该表达抑制了表达 高锌浓度。 NGO1049在人类宫颈的非政府组织感染期间高度诱导了成绩单 细胞，表明NGO1049在非政府组织发病机理中的潜在作用。根据这些发现，我假设 NGO1049是一种由ZUR调节的蛋白 上皮表面。为了检验这一假设，在此F31多样性提交中，我建议 NGO1049的定位和调节机制。其次，我将研究NGO1049对锌的贡献 在金属限制条件下采集。最后，我将定义NGO1049如何使NGO生存在发炎时 暴露于上皮细胞和含有金属螯合蛋白的人类免疫细胞后的粘膜表面。 通过定义非政府组织金属法规的新成员，我的工作有可能指向新的疗法 抗生素耐药细菌。通过细菌生理学和遗传学，生物化学和 蜂窝微生物学，以及我毕业期间可为我提供的专业发展机会 培训，这个项目将为我提供背景和专业知识，以追求职业 宿主病原体相互作用的学术研究实验室。