Nutritional regulation of pathogenesis in Staphylococcus aureus

金黄色葡萄球菌发病机制的营养调控

• 批准号: 10418664
• 负责人: Shaun R Brinsmade
• 金额: $ 40.78万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-03 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10418664
• 关键词: Affinity; Amino Acids; Anterior; Anti-Infective Agents; Antibiotics; Bacteria; Benign; Binding; Binding Sites; Biochemistry; Cells; Collaborations; Communicable Diseases; Community-Acquired Infections; Complex; Confocal Microscopy; Cues; DNase-I Footprinting; Dangerousness; Data; Developmental Process; Disease; Disease model; Electrophoresis; Endocarditis; Environment; Epithelial; Fatty Acids; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genus staphylococcus; Guanosine Triphosphate; Hospitals; Human; Immune; In Vitro; Indiana; Infection; Isoleucine; Knowledge; Leucine; Life; Life Style; Link; Logic; Mediating; Metabolic; Metabolic Pathway; Metabolism; Molecular Conformation; Molecular Genetics; Multi-Drug Resistance; N-terminal; Nutrient; Nutrient Depletion; Nutritional; Operon; Osteomyelitis; Pathogenesis; Pathogenicity; Pediatric Hospitals; Peptide Hydrolases; Peptides; Phenotype; Phosphotransferases; Physiological; Play; Pneumonia; Population; Population Density; Production; Promoter Regions; Proteins; Regulation; Regulon; Repression; Residual state; Resistance; Role; Series; Signal Transduction; Site; Skin; Skin Tissue; Soft Tissue Infections; Staphylococcal Infections; Staphylococcus aureus; Stimulus; System; Testing; Time; Toxin; Transmembrane Domain; Vaccines; Valine; Virulence; Virulence Factors; Work; alpha Toxin; antimicrobial; base; combat; commensal bacteria; design; drug resistant bacteria; genetic regulatory protein; human pathogen; in vivo; inorganic phosphate; medical schools; metabolomics; mouse model; mutant; neutrophil; novel; nutrient metabolism; overexpression; pathogen; prevent; promoter; quorum sensing; rational design; release factor; response; sensor; spreading factor; vaccine candidate

7. Project Summary/Abstract Staphylococcus aureus is the leading cause of life threatening skin and soft tissue infections, osteomyelitis, pneumonia and endocarditis, but spends much of its time as a commensal bacterium, colonizing 30-50% of the population asymptomatically. Vaccine candidates show promise for highly susceptible populations, but a general, effective approach to preventing staphylococcal infections remains elusive. Moreover, the mechanistic basis for the commensal-to-pathogen switch is complex and remains unsolved, but changing nutrient availability in the various environments of the human host likely plays an important role in informing the bacterium's decision to switch between lifestyles. The global transcriptional regulator CodY is activated by isoleucine, leucine, valine (ILV) and GTP, and adjusts metabolism and virulence gene expression. Our preliminary data indicate that CodY activity is not binary (i.e., on or off). Rather, CodY can sense a range of concentrations of ILV and GTP to generate a hierarchical transcriptional and physiological response. That is, under conditions of increasing ILV and GTP depletion, toxins and spreading factors are sequentially induced. Additional work by our lab has shown that CodY controls nearly all of the known virulence genes in conjunction with three of the most important regulators of S. aureus virulence: the Sae two-component system, the regulatory protein Rot, and the Agr quorum-sensing system. In collaboration with staphylococcal biologists Christopher Montgomery (Nationwide Children's Hospital), Victor Torres (NYU School of Medicine), and Taeok Bae (Indiana School of Medicine-NW), we will unravel the mechanistic basis by which CodY ties virulence to nutrient availability. We will use LC-MS based metabolomics, confocal microscopy, classical molecular genetics and biochemistry to (i) determine the mechanisms by which CodY regulates the expression of the sae locus, and (ii) determine how nutrient availability alters the activity of the Sae Two Component System. Additionally, we will explore mechanistically how CodY constrains virulence during S. aureus-immune cell encounters as well as during skin and soft tissue infection using a mouse model of S. aureus dermonecrosis. Understanding how nutrient signals are integrated into the virulence regulatory network by factors like CodY can potentially inform the rational design of novel antimicrobials that limit host damage, desperately replacing obsolete antibiotics in an era of pan-resistance.

7。项目摘要/摘要 金黄色葡萄球菌是威胁皮肤和软组织感染，骨髓炎的主要原因， 肺炎和心内膜炎，但大部分时间都作为共生细菌，定居30-50％ 人口无症状。候选疫苗对高度易感人群显示出希望，但 预防葡萄球菌感染的一般有效方法仍然难以捉摸。而且，机械 共同养生开关的基础是复杂的，尚未解决，但养分改变了 在人类宿主的各种环境中的可用性可能在告知 细菌决定在生活方式之间切换。全局转录调节器Cody被激活 异亮氨酸，亮氨酸，缬氨酸（ILV）和GTP，并调整代谢和毒力基因表达。我们的 初步数据表明，Cody活动不是二进制的（即打开或关闭）。相反，科迪可以感觉到一系列 ILV和GTP的浓度以产生层次转录和生理反应。那是， 在增加ILV和GTP耗竭的条件下，毒素和扩散因子被顺序诱导。 我们的实验室的其他工作表明，科迪控制了几乎所有已知的毒力基因的结合基因 带有三个最重要的金黄色葡萄球菌毒力调节剂：SAE两组分系统， 调节蛋白腐烂和AGR群体感应系统。与葡萄球菌生物学家合作 克里斯托弗·蒙哥马利（Christopher Montgomery）（全国儿童医院），Victor Torres（纽约大学医学院）和Taeok BAE（印第安纳州医学院），我们将揭开科迪将毒力与毒力联系起来的机械基础 营养可用性。我们将使用基于LC-MS的代谢组学，共聚焦显微镜，经典分子 遗传学和生物化学（i）确定Cody调节SAE表达的机制 基因座和（ii）确定养分的可用性如何改变SAE两个组件系统的活性。 此外，我们将从机械上探索Cody如何限制金黄色葡萄球菌 - 免疫细胞期间的毒力 使用金黄色葡萄球菌的小鼠模型遇到皮肤和软组织感染期间。 了解如何通过Cody等因素将营养信号集成到毒力调节网络中 可能会为新颖的抗菌剂的合理设计提供限制宿主损害的合理设计，拼命更换 在泛滥的时代过时的抗生素。

项目成果

A Fluorescence-based Method to Study Bacterial Gene Regulation in Infected Tissues.

• DOI: 10.3791/59055
• 发表时间: 2019-02
• 期刊: Journal of visualized experiments : JoVE
• 作者: R. Behera;K. D. Mlynek;Matthew S Linz;Shaun R. Brinsmade

Who's in control? Regulation of metabolism and pathogenesis in space and time.

谁在掌控？

• DOI: 10.1016/j.mib.2020.05.009
• 发表时间: 2020
• 期刊: Current opinion in microbiology
• 影响因子: 5.4
• 作者: King,AlyssaN;deMets,François;Brinsmade,ShaunR
• 通讯作者: Brinsmade,ShaunR

Regulation of the Sae Two-Component System by Branched-Chain Fatty Acids in Staphylococcus aureus.

• DOI: 10.1128/mbio.01472-22
• 发表时间: 2022-10-26
• 期刊: MBIO
• 影响因子: 6.4
• 作者: Pendleton, Augustus;Yeo, Won-Sik;Alqahtani, Shahad;DiMaggio, Dennis A., Jr.;Stone, Carl J.;Li, Zhaotao;Singh, Vineet K.;Montgomery, Christopher P.;Bae, Taeok;Brinsmade, Shaun R.
• 通讯作者: Brinsmade, Shaun R.