Regulation of the virulence factor PlcH in Pseudomonas aeruginosa

铜绿假单胞菌毒力因子PlcH的调控

基本信息

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
项目类别：
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

项目摘要

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，具有多种毒力因素，每种因素在不同的感染部位或不同的细胞类型中发挥作用。一种重要的毒力因子 Pa 是溶血性磷脂酶 C/鞘磷脂酶，PlcH。 plcH 突变体在多种方面都有缺陷感染模型和菌株的 PlcH 表达与人类疾病的严重程度和差异相关治疗效果。目前已知 PlcH 表达有三种输入：磷酸盐饥饿诱导，甘氨酸甜菜碱诱导，低氧环境抑制。虽然这些监管路径中的每一个已经描述过，我们对 PlcH 调节和改变的理解仍然存在重要差距在特定疾病背景下的这些调节途径中。在本提案中，我们将确定身份和先前未识别的 PlcH 转录和翻译后调节机制，并确定囊性纤维化患者肺部 Pa 适应过程中 PlcH 调节改变的作用。尽管PlcH对Pa毒力有贡献，但对其调控的研究落后于其他毒力因子。在这里，我们提供了 plcH 新型转录调控的初步数据，以及新的后转录调控的鉴定。翻译 PlcH 调节步骤，并呈现 plcH 直接上游代谢途径的变化囊性纤维化肺内发生的诱导。我们的假设是 PlcH 的生产和出口受到监管在多个水平上，这些监管步骤可以滴定酶产品的通量并避免对生产细胞造成损害。我们在 PlcH 方面拥有长期的经验，并拥有大型实验工具包从而接近其监管。我们有能力了解未表征背后的机制 PlcH 监管途径并确定新的监管投入。我们将通过 (i) 解决 PlcH 监管问题确定未表征的 plcH 转录调控机制以及每个机制的重要性转录诱导机制，(ii) 破译 PlcH 的翻译后控制机制，以及 (iii) 检查长期囊性纤维化过程中发生的病理适应性突变引起的 PlcH 表达变化肺部感染。这里提出的工作将使我们对 PlcH 监管有更全面的了解，提供生物学洞察并揭示抑制 PlcH 产生的潜在目标。