Eukaryotic Perception of Prokaryotic Quorum Signals

原核群体信号的真核感知

• 批准号: 7922678
• 负责人: Andrew George Palmer
• 金额: $ 5.05万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7922678
• 关键词: Affect; Alfalfa; Antibiotic Resistance; Antibiotics; Attention; Auxins; Bacteria; Biological Assay; Biological Models; Calcium; Cells; Chemicals; Clover; Combinatorial Synthesis; Complex; Death Rate; Detection; Development; Eukaryota; Exposure to; Exudate; Fluorescence; Fluorescence Microscopy; Fluorescent Probes; Gram-Negative Bacteria; HIV; Health; Hormonal; Human; Immune response; Immunocompromised Host; Individual; Infection; Ions; Laser Scanning Confocal Microscopy; Learning; Libraries; Mammalian Cell; Medicago; Methodology; Microbial Biofilms; Modeling; Nosocomial Infections; Organism; Perception; Phenotype; Physiological; Physiology; Plant Growth Regulators; Plant Model; Plant Roots; Plants; Population Density; Production; Prokaryotic Cells; Protoplasts; Pseudomonas aeruginosa; Reactive Oxygen Species; Reader; Regulation; Relative (related person); Reporter; Rhizobium radiobacter; Seedling; Signal Transduction; Surface; System; Testing; Time; Tissues; Tobacco; Trifolium repens; Virulence; Virulence Factors; Virulent; analog; antimicrobial; base; cell type; cystic fibrosis patients; design; genetic manipulation; high throughput screening; homoserine lactone; insight; mutant; novel; novel strategies; novel therapeutics; pathogen; pressure; prevent; public health relevance; quorum sensing; rapid growth; response; small molecule

DESCRIPTION (provided by applicant): Many bacteria regulate virulence factor expression as a function of their population density. This phenomenon, known as 'quorum sensing' (QS), is driven by the detection of a variety of small molecules, collectively referred to as 'autoinducers' (Als). The best characterized Als are the N-acylated L-homoserine lactones (AHLs) utilized by Gram-negative bacteria. The manipulation of AHLs to regulate virulencefactor expression, without exerting selection pressures on the pathogen, is an attractive antimicrobial strategy that has gained considerable attention. However, as the eukaryotic surfaces at which these 'chemical dialogues' between bacteria occurs are also sensitive to these signals, understanding the responses of eukaryotes to AHLs is critical to developing such novel therapeutic strategies. I propose to develop plants as a model system for defining eukaryotic responses to AHLs due to the diverse array of mutants and reporter constructs that are available, their relative ease of genetic manipulation, rapid growth rates, and well defined physiology, as well as their ability to detect and respond to AHLs. In developing this model I intend to exploit recent advances in combinatorial synthesis that have led to the development of a sizeable library of AHL analogues (" 200 compounds). This library will be screened in model plant systems like tobacco, alfalfa, and white clover to identify critical hormonal and physiological responses to a variety of structurally distinct AHLs at multiple time points. AHL responses will be evaluated by both cell-based (protoplast) assays as well as fluorescence microscopy. This two-fold approach permits both high throughput screening as well as tissue localization. Additional studies will evaluate how the perception of select AHLs affects the QS regulatory activity of the root exudate of plants. The results of these studies will be evaluated within the context of their potential impact to the development of QS-regulation as a novel strategy for regulating virulence in bacteria. PUBLIC HEALTH RELEVANCE: The majority of human infections are dependent on a complex 'chemical dialogue' exchanged between bacteria. New antimicrobial strategies, designed around the disruption of these signals, are currently under development and promise to have a substantial impact on human health. However, understanding the responses of host organisms to these bacterial signals is crucial in developing signal disruption as a novel antimicrobial strategy.

描述（由申请人提供）：许多细菌调节毒力因子的表达作为其种群密度的函数。这种现象被称为“群体感应”（QS），它是由各种小分子的检测驱动的，这些小分子统称为“自动诱导剂”（al）。最具特征的Als是革兰氏阴性菌利用的n -酰化l -同丝氨酸内酯（AHLs）。操纵ahl来调节毒力因子的表达，而不对病原体施加选择压力，是一种有吸引力的抗菌策略，已经引起了相当大的关注。然而，由于细菌之间发生这些“化学对话”的真核细胞表面也对这些信号敏感，因此了解真核细胞对ahl的反应对于开发这种新的治疗策略至关重要。我建议将植物作为一个模型系统来定义真核生物对ahl的反应，因为植物有多种可用的突变体和报告结构，它们相对容易进行遗传操作，生长速度快，明确的生理机能，以及它们检测和响应ahl的能力。在开发这个模型的过程中，我打算利用组合合成的最新进展，这些进展已经导致了一个相当大的AHL类似物库的发展（“200种化合物”）。该文库将在烟草、苜蓿和白三叶草等模式植物系统中进行筛选，以确定在多个时间点对各种结构不同的ahl的关键激素和生理反应。AHL反应将通过基于细胞（原生质体）的测定和荧光显微镜来评估。这种双重方法允许高通量筛选和组织定位。进一步的研究将评估选择ahl的感知如何影响植物根分泌物的QS调节活性。这些研究的结果将在其潜在影响的背景下进行评估，以发展qs调节作为调节细菌毒力的新策略。公共卫生相关性：大多数人类感染依赖于细菌之间复杂的“化学对话”。围绕破坏这些信号而设计的新的抗微生物战略目前正在开发中，有望对人类健康产生重大影响。然而，了解宿主生物对这些细菌信号的反应对于开发信号破坏作为一种新的抗菌策略至关重要。

项目成果

Pulse-Chase Analysis for Studies of MHC Class II Biosynthesis, Maturation, and Peptide Loading.

用于 MHC II 类生物合成、成熟和肽加载研究的脉冲追踪分析。

• DOI: 10.1007/978-1-4939-9450-2_23
• 发表时间: 2019
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Hou,Tieying;Rinderknecht,Cornelia;Ghosh,Debopam;Hadjinicolaou,AndreasV;Busch,Robert;Mellins,ElizabethD
• 通讯作者: Mellins,ElizabethD

Quorum sensing in bacterial species that use degenerate autoinducers can be tuned by using structurally identical non-native ligands.

• DOI: 10.1002/cbic.201000551
• 发表时间: 2011-01-03
• 期刊: CHEMBIOCHEM
• 影响因子: 3.2
• 作者: Palmer, Andrew G.;Streng, Evan;Jewell, Kelsea A.;Blackwell, Helen E.
• 通讯作者: Blackwell, Helen E.