Collaborative Research: Application of Transcriptomics to Understanding Mechanisms of Stress Response and Toxin Production in Microbes in Tropical Marine Waters

合作研究：应用转录组学了解热带海水微生物的应激反应和毒素产生机制

• 批准号: 1129119
• 负责人: Paul Bienfang
• 金额: $ 24.13万
• 依托单位: University of Hawaii
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2015-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1129119&HistoricalAwards=false
• 关键词: Collaborative; Research; Application; Transcriptomics; Understanding

In this project an interdisciplinary research team from the University of Hawaii at Manoa, Stanford University, and Rutgers University will study the transcriptomes of Gambierdiscus, Staphylococcus aureus, and Enterococcus in Hawaiian coastal waters - toxin-producing species that pose significant health hazards to humans. Their goal is to elucidate the mechanisms associated with toxin production by Gambierdiscus and inactivation by sunlight of the bacterial pathogen, S. aureus, and the fecal indicator, Enterococcus. Because the transcriptome of an organism reflects the genes that are actively being expressed at any given time, this study, by elucidating levels of messenger RNA expression in the target organisms, should provide valuable insights into the mechanisms responsible for toxin production, responses to stress and/or subsequent bacterial inactivation. The research program will be organized around four working hypotheses: H1: Production of ciguatoxin by Gambierdiscus reflects the physiological condition of the culture as influenced by temperature, irradiance, and growth phase. H2: High throughput transcriptome analysis from toxin (+) and toxin (-) cultures will allow the identification of genes involved in ciguatoxin production and understanding of the light and nutrient conditions that favor this function. H3: Whereas E. faecalis is susceptible to indirect photoinactivation under exposure to sunlight in seawater, S. aureus is not. This is likely due to the presence of carotenoid pigments within the cells, which are able to quench reactive oxygen species. H4: E. faecalis and S. aureus respond to photo-stress in clear seawater by up-regulating genes that encode for proteins to repair cellular damage and mitigate oxidative stress; this "stressome" will change with increasing exposure to photostress, revealing the stress at which repair is no longer possible. Broader Impacts: The information that will be gathered in this study is expected to contribute significantly to advancing the field of risk management with respect to recreational water use and seafood consumption beyond reliance on empirical correlations and towards policies that are based on a mechanistic understanding of the threats these organisms pose to human health.

在这个项目中，来自夏威夷大学，斯坦福大学和罗格斯大学的跨学科研究小组将研究夏威夷沿海水域的gambierdiscus，金黄色葡萄球菌和肠球菌的转录组 - 生产巨大的健康危险性，使夏威夷沿海水域 - 毒素产生了大量的健康危害。他们的目标是阐明甘比尔氏菌与毒素产生相关的机制，并通过细菌病原体的金黄色葡萄球菌和粪便指示剂肠球菌灭活。由于生物体的转录组反映了在任何给定时间积极表达的基因，因此通过阐明目标生物中的使者RNA表达水平的水平，应提供对负责毒素产生的机制的有价值的见解，对压力的反应和/或随后的细菌失活。该研究计划将围绕四个工作假设组织：H1：GambierDiscus生产Ciguatoxin，反映了受温度，辐照度和生长阶段影响的培养物的生理状况。 H2：来自毒素（+）和毒素（ - ）培养物的高吞吐量转录组分析将允许鉴定涉及雪茄毒素产生的基因以及对有利于这种功能的光和营养条件的理解。 H3：虽然E.粪便易于在海水暴露于阳光下间接光活化，但金黄色葡萄球菌却没有。这可能是由于细胞内存在类胡萝卜素色素，能够淬灭活性氧。 H4：E. faecalis和S.金黄色葡萄球菌通过上调蛋白质来修复细胞损伤并减轻氧化应激的基因来应对透明海水中的照相压力；这种“压力”将随着照相金的曝光而变化，揭示了不再可能修复的压力。更广泛的影响：预计将在这项研究中收集的信息将在娱乐性用水和海鲜消耗方面促进风险管理领域，而不仅仅是基于对威胁的机械理解这些生物对人类健康构成威胁的机械理解的政策。