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.

摘要 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。由于淋病发病率的增加和抗生素耐药性的增加，疫苗和新的 迫切需要治疗方法。一个有前途的战略是针对营养金属收购系统，因为他们 通常是保守的，在感染期间表达，并且是细菌在体内存活所必需的。Ngo破坏了 宿主金属限制机制通过表达外膜转运蛋白来获取必需金属 人金属螯合蛋白，例如来自钙卫蛋白和银屑病蛋白的锌和来自乳铁蛋白的铁， 转铁蛋白然而，许多支持非政府组织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，在生长于锌环境下的Ngo中，ngo 1049基因的产物高度表达。 限制条件。Ngo 1049在致病性奈瑟菌中是保守的，生物信息学分析预测， Ngo 1049是一种定位于周质中的金属结合转移酶。在上游发现了一个Zur结合基序 表明表达受Zur（锌吸收调节剂）调节，Zur抑制了ngo 1049的表达。 高锌浓度。在人宫颈内膜Ngo感染过程中，ngo 1049转录物被高度诱导 细胞，表明Ngo 1049在Ngo发病机制中的潜在作用。基于这些发现，我假设 Ngo 1049是一种可调节的蛋白质，可促进炎症时金属限制条件下锌的获取。 上皮表面为了验证这一假设，在本F31多样性提交文件中，我建议确定 Ngo 1049的定位和调节机制。其次，我将研究Ngo 1049对锌的贡献。 在金属限制条件下获得。最后，我将定义Ngo 1049如何使Ngo在发炎中存活 在暴露于含有金属螯合蛋白的上皮细胞和人免疫细胞后，粘膜表面的金属螯合蛋白质。 通过定义Ngo金属调节子的这个新成员，我的工作可能会指向新的治疗方法。 耐药细菌。通过细菌生理学和遗传学、生物化学的结合， 细胞微生物学，以及在我毕业期间提供给我的专业发展机会 培训，这个项目将为我提供背景和专业知识，以追求职业生涯的领导者， 在宿主-病原体相互作用的学术研究实验室。