Vibrio cholerae biofilms: structure, function, regulation and role in infection

霍乱弧菌生物膜：结构、功能、调节和在感染中的作用

• 批准号: 9507758
• 负责人: Havva Fitnat Yildiz
• 金额: $ 44.14万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9507758
• 关键词: Address; Architecture; Area; Binding; Biology; Cells; Cholera; Clinical; Communities; Couples; Developing Countries; Development; Disasters; Disease; Disease Outbreaks; Ecosystem; Environment; Environmental Risk Factor; Epidemic; Extracellular Matrix; Family; Future; Gene Cluster; Genes; Genetic Transcription; Guanosine Monophosphate; Habitats; Human; Image; Industrialization; Infection; Intestines; Life Cycle Stages; Location; Maintenance; Mediating; Microbe; Microbial Biofilms; Microscopy; Modification; Molecular; Patients; Periodicity; Phosphorylation; Polysaccharides; Process; Production; Proteins; Proteolysis; Regulation; Regulon; Resolution; Role; Structure; Surface; Testing; Transcriptional Regulation; Variant; Vibrio; Vibrio cholerae; base; combat; genetic regulatory protein; improved; interfacial; man; mechanical properties; member; microbial community; new therapeutic target; pathogen; public health relevance; sensor histidine kinase; stool sample; tool development; transmission process

DESCRIPTION (provided by applicant): Vibrio cholerae causes the disease cholera and is a natural inhabitant of aquatic environments. Seasonal cholera outbreaks occur where the disease is endemic and can spread worldwide. V. cholerae's ability to form biofilms (i.e., matrix-enclosed, surface-associated communities) is crucial for its survival in aquatic habitats between epidemics and is advantageous for host-to-host transmission during epidemics. The objective of this proposal is to improve our understanding of biofilm matrix components, the mechanisms and regulation of biofilm formation, and their importance in the biology of V. cholerae. In Aim 1, we will focus on understanding how biofilm matrix proteins RbmA, RbmC and Bap1 function. We will determine the molecular basis of RbmA and VPS interactions and determine molecular determinants and consequences of RbmA proteolysis. We will test whether RbmC and Bap1 bind to VPS and identify regions important for their function. We will also determine how these proteins contribute to the mechanical properties of biofilms, as well as the location of each matrix component by super resolution microscopy. In Aim 2, we will determine the mechanism used by the regulatory proteins, VpsR and VpsT, in biofilm formation. We will identify sensor histidine kinases that phosphorylate VpsR, and determine phosphorylation dynamics of VpsR during biofilm development. We will also identify direct targets of VpsT and VpsR during biofilm formation and ascertain the contribution of VpsR and VpsT regulon members in V. cholerae transmission and dissemination into the environment. Clarifying the mechanisms of biofilm formation in V. cholerae will prove useful for the development of future strategies for controlling cholera epidemics, and facilitate identification of novel drug targets for combating the pathogen during infection.

描述（由申请人提供）：霍乱弧菌引起霍乱疾病，是水生环境的天然居民。季节性霍乱爆发发生在疾病流行的地方，并且可以在世界范围内传播。霍乱弧菌形成生物膜（即基质封闭、表面相关群落）的能力对于其在流行病期间在水生栖息地中的生存至关重要，并且有利于流行病期间的宿主间传播。该提案的目的是提高我们对生物膜基质成分、生物膜形成的机制和调节及其在霍乱弧菌生物学中的重要性的理解。在目标 1 中，我们将重点了解生物膜基质蛋白 RbmA、RbmC 和 Bap1 的功能。我们将确定 RbmA 和 VPS 相互作用的分子基础，并确定 RbmA 蛋白水解的分子决定因素和后果。我们将测试 RbmC 和 Bap1 是否与 VPS 结合并确定对其功能重要的区域。我们还将通过超分辨率显微镜确定这些蛋白质如何影响生物膜的机械特性，以及每个基质成分的位置。在目标 2 中，我们将确定调节蛋白 VpsR 和 VpsT 在生物膜形成中使用的机制。我们将鉴定磷酸化 VpsR 的传感器组氨酸激酶，并确定生物膜形成过程中 VpsR 的磷酸化动态。我们还将确定生物膜形成过程中 VpsT 和 VpsR 的直接靶标，并确定 VpsR 和 VpsT 调节子成员在霍乱弧菌传播和传播到环境中的贡献。阐明霍乱弧菌生物膜形成的机制将有助于制定未来的控制策略 霍乱流行，并促进识别在感染期间对抗病原体的新药物靶点。