Regulation of the virulence factor PlcH in Pseudomonas aeruginosa

铜绿假单胞菌毒力因子PlcH的调控

• 批准号: 10884588
• 负责人: MATTHEW J WARGO
• 金额: $ 37.88万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-18 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10884588
• 关键词: Betaine; Biological; Cells; Choline; Cystic Fibrosis; Data; Disease; Environment; Epithelial Cells; Evolution; Functional disorder; Gene Expression Regulation; Genetic; Genetic Transcription; Glycine; Health; Homeostasis; Human; Individual; Infection; Iron; Knowledge; Left; Lung; Lung infections; Metabolic Pathway; Metabolism; Miltefosine; Mission; Modeling; Mus; Mutation; Oxygen; Pathogenesis; Pathway interactions; Persons; Phospholipase C; Phospholipids; Pilum; Play; Positioning Attribute; Post-Translational Regulation; Process; Production; Pseudomonas aeruginosa; Pulmonary Cystic Fibrosis; Pulmonary Surfactants; Regulation; Regulatory Pathway; Repression; Research; Role; Severity of illness; Signal Transduction; Site; Sphingolipids; Sphingomyelinase; Sphingosine; Starvation; System; Testing; Titrations; Transcriptional Regulation; Treatment Efficacy; Type III Secretion System Pathway; Type IV Secretion System Pathway; United States National Institutes of Health; Virulence; Virulence Factors; Work; biochemical tools; cell type; detection platform; dimethylglycine; drug repurposing; experience; extracellular; human disease; human pathogen; inorganic phosphate; insight; mutant; novel; pathogen; periplasm; surfactant; targeted treatment; therapeutic target; tissue culture

Pseudomonas aeruginosa (Pa) is an important opportunistic human pathogen that deploys a variety of virulence factors, each playing roles in different infection sites or with different cell types. One important virulence factor of Pa is the hemolytic phospholipase C/sphingomyelinase, PlcH. plcH mutants are defective in a wide range of infection models and PlcH expression by strains has been correlated to human disease severity and differential treatment efficacy. There are three currently known inputs to PlcH expression: induction by phosphate starvation, induction by glycine betaine, and repression by low oxygen environments. While each of these regulatory paths have been described, there remain important gaps in our understanding of PlcH regulation as well as alterations in these regulatory pathways in specific disease contexts. In this proposal, we will determine the identity and mechanisms of previously unidentified transcriptional and posttranslational regulation of PlcH and determine the role of altered PlcH regulation during Pa adaptation in the lungs of people with cystic fibrosis. Despite the contribution of PlcH to Pa virulence, study of its regulation lags behind that of other virulence factors. Here we present preliminary data for novel transcriptional regulation of plcH, identification of a new post- translational PlcH regulatory step, and present changes to metabolic pathways directly upstream of plcH induction that occur within the cystic fibrosis lung. Our hypothesis is that PlcH production and export is regulated at multiple levels and that these regulatory steps work to titrate the flux of enzymatic products and to avoid damage to producing cells. We have longstanding experience with PlcH and a large experimental toolkit with which to approach its regulation. We are well positioned to understand the mechanisms behind uncharacterized PlcH regulatory pathways and to identify new regulatory inputs. We will address PlcH regulation by (i) determining the mechanisms of uncharacterized plcH transcriptional regulation and the importance of each mechanism of transcriptional induction, (ii) deciphering post-translational control mechanisms for PlcH, and (iii) examining alteration in PlcH expression by pathoadaptive mutations occurring during long term cystic fibrosis lung infections. The work proposed here will allow us to have a more complete understanding of PlcH regulation, providing biological insight and exposing potential targets for inhibition of PlcH production.

铜绿假单胞菌（PA）是一种重要的机会性人类病原体，可部署多种毒力 因素，每个在不同的感染部位或不同细胞类型中扮演角色。一个重要的毒力因子 PA的是溶血磷脂酶C/鞘磷脂酶，Plch。 PLCH突变体在广泛的范围内有缺陷 感染模型和菌株的表达与人类疾病的严重程度和差异有关 治疗功效。当前有三个已知的PLCH表达输入：通过磷酸盐饥饿诱导， 甘氨酸甜菜碱的诱导和低氧环境的抑制作用。而这些监管路径中的每一个 已经描述了我们对PLCH调节的理解以及更改的重要差距 在特定疾病环境中的这些调节途径中。在此提案中，我们将确定身份和 PLCH先前未识别的转录和翻译后调节的机制，并确定 囊性纤维化患者肺中PA适应过程中PLCH调节改变的作用。 尽管PLCH对PA毒力有贡献，但研究其调节落后于其他毒力因素。 在这里，我们介绍了用于新的PLCH转录调节的初步数据，鉴定了新的后 转化PLCH调节步骤，并在PLCH上游直接上游代谢途径进行更改 囊性纤维化肺中发生的诱导。我们的假设是调节PLCH的生产和出口 在多个层面上，这些调节步骤可滴定酶促产品的通量，并避免 损害产生细胞。我们在PLCH和大型实验工具包中有长期的经验 这要处理其法规。我们很适合了解未表征背后的机制 PLCH监管途径并确定新的监管输入。我们将通过（i）解决PLCH法规 确定未表征的PLCH转录调节的机制和每个的重要性 转录诱导的机制，（ii）PLCH的解密后翻译后控制机制和（iii） 通过长期囊性纤维化期间发生的病原体突变检查PLCH表达的改变 肺部感染。 这里提出的工作将使我们对PLCH法规有更全面的了解 生物学洞察力并暴露了抑制PLCH生产的潜在靶标。