Vibrio Cholerae Colonization of Chitin Surfaces

霍乱弧菌在甲壳质表面的定植

• 批准号: 7214157
• 负责人: Gary K Schoolnik
• 金额: $ 33.57万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2009-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7214157
• 关键词: Acetylglucosamine; Affect; Bacteria; Carbon; Cells; Chemotaxis; Chitin; Chitinase; Cholera; Collaborations; Communities; Condition; Copepoda; Data; Defect; Development; Disease; DsRed; Ecosystem; Epidemic; Gene Expression; Genes; Genetic; Genomics; Gills; Green Fluorescent Proteins; Growth; Habitats; Human; Image; In Situ; Indigenous; Individual; Infectious Agent; Laser Scanning Confocal Microscopy; Libraries; Marines; Mediating; Membrane; Methods; Microbial Biofilms; Microscopy; Molecular Profiling; Montana; Movement; Mutation; Nitrogen; Nutrient; Phase; Phenotype; Polymers; Polysaccharides; Population; Process; Proteins; Rate; Regulon; Research Project Grants; Role; Simulate; Source; Surface; Swimming; System; Universities; Vibrio cholerae; Vibrio cholerae O1; Work; Zooplankton; cell motility; cellular imaging; exoskeleton; extracellular; mutant; pathogen; professor; programs; promoter; research study; sensor histidine kinase

DESCRIPTION (provided by applicant): Vibrio cholerae O1, a cause of epidemic diarrheal disease, normally resides as an indigenous component of riverine, estuarine and marine ecosystems. In these habitats, it associates with the chitinous exoskeletons of zooplankton. The principal objective of this research project is to characterize the interaction of this human pathogen with a chitin surface. Chitin is an insoluble polymer of N-acetylglucosamine; through the secretion of chitinases, V. cholerae can use chitin as a sole source of carbon and nitrogen in nutrient-poor aquatic habitats. This project will explore the hypothesis that chitin utilization by V. cholerae is a four step process: chemotaxis toward and attachment to the chitin surface; horizontal dispersal of attached bacteria across the surface; vertical growth of the attached population as a biofilm community; and detachment from the biofilm surface and resumption of the planktonic mode-of-growth. The work proposed here will explore each of these hypothesized steps through the combined use of genomic, genetic and cell imaging methods. In collaboration with Professor Gill Geesey at Montana State University, reflected differential interference contrast (DIC) and epifluorescence microscopy will be used to image individual cells as they attach to and spread across a synthetic chitin membrane attached to a laminar flow cell experimental system. Scanning confocal laser microscopy (SCLM) will be employed at the Stanford Biofilm Center to capture the temporal and spatial features of biofilm development on the chitin surface. Microarray expression profiling will be used to identify genes, which comprise a hypothesized detachment regulon. Studies undertaken in collaboration with Professor Saul Roseman at Johns Hopkins University will examine the role of a newly identified chitin sensor histidine kinase protein in each of the four chitin utilization steps. Green fluorescent protein (GFP) promoter fusions will be used to disclose when and where individual genes are expressed on the chitin surface and mutants will be sought that are defective in the chitin utilization program. In the last Specific Aim of the proposed work, these results will be examined using experimental systems that more closely resemble conditions in natural aquatic ecosystems including their relevance for the colonization of swimming copepods. If successful, this project should generate new information about the persistence and control of infectious agents in aquatic reservoirs.

描述（由申请人提供）：流行腹泻病的原因纤维状霍乱O1通常是河流，河口和海洋生态系统的土著成分。在这些栖息地中，它与浮游动物的奇特的外骨骼相关联。该研究项目的主要目标是表征这种人类病原体与几丁质表面的相互作用。几丁质是N-乙酰葡萄糖的不溶性聚合物。通过几丁质酶的分泌，V。Cholerae可以在营养贫困的水生栖息地中使用几丁质作为唯一的碳和氮来源。该项目将探讨以下假设：V。Cholerae的几丁质利用是一个四步骤过程：趋化和固定在几丁质表面上；整个表面上附着的细菌的水平分散；作为生物膜社区，附着的人群的垂直增长；并脱离生物膜表面，并恢复浮游性增长方式。这里提出的工作将通过基因组，遗传和细胞成像方法的结合使用来探索这些假设的步骤。与蒙大拿州立大学的吉尔·盖西（Gill Geesey）教授合作，反射的差异干扰对比度（DIC）和落荧光显微镜将用于对单个细胞进行图像图像，因为它们附着并散布在附着于层状流量流细胞实验系统上的合成几根膜上。扫描共聚焦激光显微镜（SCLM）将在斯坦福生物膜中心使用，以捕获几丁质表面生物膜发育的时间和空间特征。微阵列表达分析将用于鉴定基因，该基因包括假设的分离调节。与约翰·霍普金斯大学（Johns Hopkins University）的索尔·罗斯曼（Saul Roseman）教授合作进行的研究将研究新近鉴定的几丁质传感器组氨酸激酶蛋白在四个几丁质利用步骤中的每个步骤中的作用。绿色荧光蛋白（GFP）启动子融合将用于披露在几丁质表面上表达单个基因的何时何地，并且将寻求在甲壳质利用率程序中有缺陷的突变体。在拟议的工作的最后一个具体目的中，将使用实验系统对这些结果进行检查，这些实验系统更像天然水生生态系统中的条件，包括它们与游泳copepods定殖的相关性。如果成功，该项目应生成有关水库中传染剂的持久性和控制的新信息。

项目成果

RpoS controls the Vibrio cholerae mucosal escape response.

• DOI: 10.1371/journal.ppat.0020109
• 发表时间: 2006-10
• 期刊: PLOS PATHOGENS
• 影响因子: 6.7
• 作者: Nielsen, Alex Toftgaard;Dolganov, Nadia A;Otto, Glen;Miller, Michael C;Wu, Cheng Yen;Schoolnik, Gary K
• 通讯作者: Schoolnik, Gary K

Serogroup conversion of Vibrio cholerae in aquatic reservoirs.

• DOI: 10.1371/journal.ppat.0030081
• 发表时间: 2007-06
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Blokesch M;Schoolnik GK
• 通讯作者: Schoolnik GK

A bistable switch and anatomical site control Vibrio cholerae virulence gene expression in the intestine.

• DOI: 10.1371/journal.ppat.1001102
• 发表时间: 2010-09-16
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Nielsen AT;Dolganov NA;Rasmussen T;Otto G;Miller MC;Felt SA;Torreilles S;Schoolnik GK
• 通讯作者: Schoolnik GK