Form and Function of Tandem Duplicate Small RNA Molecules in Shigella dysenteriae

痢疾志贺氏菌串联重复小 RNA 分子的形式和功能

基本信息

批准号：
8574164
负责人：
ERIN R MURPHY
金额：
$ 44.55万
依托单位：
OHIO UNIVERSITY ATHENS
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-16 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8574164
关键词：
Achievement Antibiotic Resistance Antibiotics Attention Bacteria Biological Assay Cell division Communicable Diseases Computer Simulation Data Development Disease Dysentery Environment Equilibrium Foundations Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Transcription Goals Human Human body Incidence Infection Infection prevention Invaded Iron Knowledge Mediating Mentors Modeling Molecular Nutrient Pathogenesis Pathway interactions Pattern Phenotype Physiology Play Positioning Attribute Process Production RNA Regimen Regulation Research Role Shigella Shigella Infections Shigella boydii Shigella dysenteriae Shigella flexneri Small RNA Structure Students System Temperature Testing Therapeutic Training Training Programs Vaccines Virulence Water antimicrobial drug bacterial genetics base commensal microbes experience human disease innovation model development novel therapeutics pathogen pathogenic bacteria prevent public health relevance response skills

项目摘要

DESCRIPTION (provided by applicant): Shigellosis, a several diarrheal disease caused by infection with bacteria of the genus Shigella, remains endemic throughout the world. The global burden of shigellosis is due in part to the lack of a vaccine to prevent the infection and the lack of a universally safe and available antibiotic regimen to treat the infection. While the potential f using small RNA molecules (sRNAs) as models for, or targets of, RNA-based anti-Shigella therapeutics is promising, what is lacking is a comprehensive understanding of the role that sRNAs play in controlling Shigella physiology and pathogenesis. A necessary first step towards achieving the long-term goal of developing RNA-based antibiotics to treat shigellosis is to reveal the full extent to which sRNAs control the physiology and virulence of Shigella species. To this end, the overall objective of this study is to elucidate the role of newly identified duplicate sRNAs RyfA1 and RyfA2 in controlling the physiology and pathogenesis of S. dysenteriae. The central hypothesis being tested is that RyfA1 and RyfA2 are differentially produced under unique environmental conditions and modulate S. dysenteriae virulence by regulating the expression of distinct over-lapping sets of genes. This hypothesis will be tested by achieving the following specific aims: 1) identify the iron- and temperature-responsive factors regulating the production of RyfA1 and RyfA2; 2) elucidate the function of RyfA1 and RyfA2 by identifying the regulatory targets of each; and 3) determine the effect of RyfA1 and RyfA2, individually and in combination, on S. dysenteriae virulence. The proposed systematic characterization of duplicate sRNAs, RyfA1 and RyfA2, will contribute to answering fundamental questions in the fields of Shigella pathogenesis and bacterial sRNAs, contributions that will facilitate achievement of the long-term goal of developing therapeutics to treat shigellosis. The proposed study puts forth the innovative hypothesis that two sRNA molecules that share 95% sequence identity and a identical predicted structure have non-redundant functions. This hypothesis will be tested using a balanced approach of standard, student friendly, bacterial genetics and innovative high-tech assays. Student involvement in every step of the proposed study form the foundation of an innovative training program that will provide students with practical experience in the fields of bacterial pathogenesis and RNA-based regulation. The resulting combination of skills and experience will position students well to contribute to the rapidly advancing fields of ribo-regulation in pathogenic bacteria, commensal bacteria and eukaryotic systems.

描述（由申请人提供）：志贺氏菌病是一种由志贺氏菌属细菌感染引起的几种腹泻疾病，在世界各地仍然流行。志贺氏菌病的全球负担部分是由于缺乏预防感染的疫苗和缺乏一种普遍安全且可用的抗生素疗法来治疗感染。虽然使用小 RNA 分子 (sRNA) 作为基于 RNA 的抗志贺氏菌疗法的模型或靶点的潜力是有希望的，但缺乏的是对 sRNA 在控制志贺氏菌生理学和发病机制中所发挥的作用的全面了解。实现开发基于 RNA 的抗生素来治疗志贺氏菌病的长期目标，必要的第一步是揭示 sRNA 控制志贺氏菌属的生理和毒力的全部程度。为此，本研究的总体目标是阐明新鉴定的重复 sRNA RyfA1 和 RyfA2 在控制痢疾沙门氏菌生理和发病机制中的作用。正在测试的中心假设是，RyfA1 和 RyfA2 在独特的环境条件下差异产生，并通过调节不同重叠基因组的表达来调节痢疾沙门氏菌毒力。该假设将通过实现以下具体目标进行检验：1）确定调节 RyfA1 和 RyfA2 产生的铁和温度响应因子； 2) 通过确定RyfA1和RyfA2的调控靶点来阐明RyfA1和RyfA2的功能； 3)确定RyfA1和RyfA2单独和组合对痢疾沙门氏菌毒力的影响。拟议的重复 sRNA RyfA1 和 RyfA2 的系统表征将有助于回答志贺氏菌发病机制和细菌 sRNA 领域的基本问题，有助于实现开发治疗志贺氏菌病的疗法的长期目标。该研究提出了一个创新假设，即两个具有 95% 序列同一性和相同预测结构的 sRNA 分子具有非冗余功能。该假设将使用标准、学生友好的细菌遗传学和创新高科技检测的平衡方法进行测试。学生对拟议研究每一步的参与构成了创新培训计划的基础，该计划将为学生提供细菌发病机制和基于 RNA 的调控领域的实践经验。由此产生的技能和经验的结合将使学生能够为病原菌、共生细菌和真核系统中快速发展的核糖调节领域做出贡献。