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

环境条件的变化控制着细菌小非编码RNA(SRNAs)的表达，sRNAs调节其他基因的表达。虽然sRNA在调节细菌基因表达和生理方面发挥着重要作用，但要了解它们的功能仍然存在重大障碍。该项目结合了遗传学、生理学和转录学的方法来识别sRNA基因，并阐明这些sRNA的表达如何影响细菌细胞生理学。这项工作是在埃尼希瓦氏菌中进行的，这是一种具有利用金属代替氧气进行呼吸的耐人寻味的细菌。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展示他们的研究成果。这位首席调查员和他的学生通过指导当地中小学生进行科学项目来参与社区推广。该项目将调查高度保守的RNA伴侣分子Hfq在调节S.onedensis新陈代谢和应激反应中的sRNA信号中的作用。这项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在一起。RNA-Seq用于识别在目的条件下表达的假定sRNA。使用遗传功能丧失和功能获得的方法操纵sRNA表达将被用来测试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。鉴于细菌中存在大量的sRNA基因，本研究着重于sRNA的功能，该功能可能与圆顶螺旋藻的金属还原代谢有关。研究成果将通过同行评议的期刊出版物以及在区域、国家和国际会议上的陈述进行传播。

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.