RUI: Hfq and sRNA Function in Shewanella oneidensis Growth and Adaptation to Stress

RUI：Hfq 和 sRNA 在 Shewanella oneidensis 生长和应激适应中的功能

• 批准号: 1456536
• 负责人: Brett Pellock
• 金额: $ 20.09万
• 依托单位: Providence College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1456536&HistoricalAwards=false
• 关键词: RUI; Hfq; sRNA; Function; Shewanella

Changes in environmental conditions control the expression of bacterial small noncoding RNAs (sRNAs) that regulate the expression of other genes. Though sRNAs play important roles in adjusting bacterial gene expression and physiology, there are significant hurdles to understanding their function. This project combines genetic, physiological, and transcriptomic approaches to identify sRNA genes and elucidate how expression of these sRNAs impacts bacterial cell physiology. This work is being performed in Shewanella oneidensis, a bacterium with the intriguing capacity to respire using metals in the place of oxygen. Students participate fully in all aspects of the study, including experimental design, performing experiments, data analyses, and manuscript preparation and revision. Students also regularly present their work at scientific conferences. The principle investigator and his students participate in community outreach by mentoring local primary and secondary school students performing science projects.The project will investigate the role of a highly-conserved RNA chaperone molecule, Hfq, in regulation of sRNA signaling in S. oneidensis metabolism and stress responses. The research couples discovery-based systematics (next generation RNA sequencing) with molecular, genetic, and physiological approaches. RNA-Seq is used to identify putative sRNAs expressed under conditions of interest. Manipulating sRNA expression using genetic loss-of-function and gain-of-function approaches will be used to test whether sRNAs regulate predicted mRNA targets and to determine how sRNA expression impacts cell physiology. Given the large number of sRNA genes in bacteria, this study focuses on sRNA functions likely to be linked to the metal reducing metabolism of S. oneidensis. Results from the research effort will be disseminated through peer-reviewed journal publications, and through presentations at regional, national, and international meetings.

环境条件的变化控制细菌小的非编码RNA（sRNA）的表达，这些小的非编码RNA调节其他基因的表达。虽然sRNAs在调节细菌基因表达和生理方面发挥着重要作用，但了解它们的功能仍存在重大障碍。该项目结合了遗传学，生理学和转录组学方法来鉴定sRNA基因，并阐明这些sRNA的表达如何影响细菌细胞生理学。这项工作正在希瓦氏菌中进行，希瓦氏菌是一种具有利用金属代替氧气进行呼吸的有趣能力的细菌。学生充分参与研究的各个方面，包括实验设计，执行实验，数据分析，以及手稿的准备和修改。学生们还定期在科学会议上介绍他们的工作。 主要研究者和他的学生参与社区外展，指导当地的小学和中学学生进行科学项目。该项目将研究高度保守的RNA伴侣分子Hfq在S.代谢和应激反应。该研究将基于发现的系统学（下一代RNA测序）与分子，遗传和生理方法结合起来。RNA-Seq用于鉴定在感兴趣的条件下表达的推定sRNA。使用遗传功能丧失和功能获得方法操纵sRNA表达将用于测试sRNA是否调节预测的mRNA靶标，并确定sRNA表达如何影响细胞生理学。 鉴于细菌中大量的sRNA基因，本研究的重点是sRNA的功能可能与金属还原代谢的S。oneidensis。 研究工作的结果将通过同行评审的期刊出版物以及在区域、国家和国际会议上的演讲进行传播。