Biogenesis and Function of Bacterial Outer Membrane Vesicles

描述（由申请人提供）：该项目的长期目标是了解革兰氏阴性菌产生的外膜囊泡的基本方面：它们是如何产生的？外膜囊泡占生长培养物中总外膜蛋白的0.1%至1%，并且由迄今为止研究的所有非致病性和致病性细菌产生，其中毒性细菌通常产生毒性囊泡并且比非致病性对应物产生更多。然而，关于它们如何以及为什么被制造的基本过程是未知的。以前，我们已经发现外膜囊泡有助于革兰氏阴性菌的基本应激反应。我们建议，外膜囊泡的生产是由特定的信封组件，并使重塑的周质和外膜内容，以应对环境的变化和压力。细胞被膜蛋白和脂质的快速替换以及囊泡介导的有毒组分的释放可能是细菌在环境中存活的关键因素，特别是为了适应压力。利用本实验室新发现的囊泡形成突变体和生物化学技术，本研究旨在阐明囊泡有助于细菌适应的机制。本研究的研究目标如下：1）表征细菌囊泡形成被膜对应激的响应，2）鉴定和表征囊泡形成途径中的基因，3）表征囊泡在细胞被膜重塑中的作用。这些结果将产生期待已久的关于囊泡形成所需的细菌过程的见解，并将揭示关于这种新描述的应激反应的细节。压力产物管理的一般生物学原理将通过比较我们在细菌研究中的发现与相关的真核外泌体来揭示。对人类健康的影响是表征一种常见的细菌过程，可以在治疗上靶向该过程，以增加细菌对抗生素治疗的敏感性。 公共卫生相关性：该项目通过详细介绍我们对细菌囊泡形成的理解而有益于公众健康，这是一种分泌过程，常见于太多的致病细菌，使细菌能够抵抗抗生素并将有毒因子分散到受感染的组织中。拟议的研究重点是揭示囊泡形成的分子基础，旨在促进这一潜在治疗靶点的开发。

DESCRIPTION (provided by applicant): The long-term goal of this project is to understand the fundamental aspects of outer membrane vesicles produced by Gram-negative bacteria: How and why are they made? Outer membrane vesicles represent 0.1 to 1% of total outer membrane protein in a growing culture and are produced by all non-pathogenic and pathogenic bacteria studied to date, with virulent bacteria typically producing toxic vesicles and more than non-pathogenic counterparts. Yet the basic processes regarding how and why they are made are unknown. Previously, we have found that outer membrane vesicles contribute a basic stress response of Gram-negative bacteria. We propose that outer membrane vesicle production is governed by specific envelope components and enables remodeling of the periplasmic and outer membrane content in response to environmental changes and stresses. Rapid replacement of cell envelope protein and lipid and vesicle-mediated release of toxic components could be key elements of bacterial survival in the environment, particularly to accommodate stress. Using newly identified vesiculation mutants and biochemistry techniques developed in this laboratory, this research aims to elucidate the mechanism by which vesicles contribute to bacterial adaptation. The research objectives of this study are the following: 1) Characterize the bacterial vesiculation envelope response to stress, 2) Identify and characterize genes in the vesiculation pathway, and 3) Characterize the role of vesicles in cell envelope remodeling. The results will yield long-awaited insight regarding the bacterial processes required for vesicle formation and will reveal details regarding this newly describe stress response. General biological principles of stress product management will be revealed by comparing our findings in the study of bacteria to the related eukaryotic exosomes. The impact on human health is the characterization of a common bacterial process that can be targeted therapeutically to increase bacterial sensitivity to antibiotic treatment. PUBLIC HEALTH RELEVANCE: This project benefits public health by detailing our understanding of bacterial vesiculation, a secretory process common too many pathogenic bacteria that enables bacteria to resist antibiotics and to disperse toxic factors into infected tissues. The proposed research focuses on exposing the molecular basis for vesiculation and aims to facilitate development of this potential therapeutic target.