Molecular mechanisms underlying the regulation of gene expression by non-coding RNAs in archaea

古细菌非编码RNA调控基因表达的分子机制

• 批准号: 2223488
• 负责人: Jocelyne DiRuggiero
• 金额: $ 71.3万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2025-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2223488&HistoricalAwards=false
• 关键词: Molecular; mechanisms; underlying; regulation; gene

This project investigates the role of small non-coding RNAs (sRNAs) in the response of microorganisms to environmental stress. Hundreds of sRNAs have been discovered in Archaea, a group of microorganisms with a close evolutionary relationship with Eukaryotes, but little is known about their mechanisms of action and how they modulate the functions of living organisms through evolution. The research will elucidate the regulatory landscape of an sRNA in a model archaeon in order to address this gap in knowledge. A deeper understanding of archaeal sRNA functions and regulation will enable manipulation of sRNA molecules as synthetic biology tools and rational design of biological systems for biotechnology applications. The broader impact activities will engage students from diverse backgrounds early in their STEM careers with laboratory research, especially first-generation college students, women, and underrepresented minorities. Students will be recruited through FiGURE (First Generation Undergrads Research Experience), an innovative research experience and cohort building program at Johns Hopkins University. These activities will also provide mentorship training for graduate students.This research aims to gain a mechanistic understanding of sRNA regulation in a model archaeon, Haloferax volcanii (Hv) in response to oxidative stress. The specific goals are to (1) characterize post-transcriptional mechanisms of the SHOxi sRNA-mediated oxidative stress response and determine how SHOxi regulates putative targets by modulating mRNA stability or interfering with translation; (2) identify chaperone proteins and other factors involved in the interactions of oxidative stress-responsive sRNAs with their targets; and (3) elucidate how the stress-responsive sRNA SHOxi is regulated under oxidative stress (particularly testing the hypothesis that canonical archaeal transcription factors regulate SHOxi). The research will employ a combination of genetic and biochemical approaches, site-directed mutagenesis combined with sRNA-mRNA interaction modeling, and high-throughput phenotyping. The outcomes are expected to reveal mechanisms by which an archaeal sRNA binds and regulates its targets, identify novel RNA binding proteins, and discover how sRNA-mediated regulatory networks are integrated with those of transcription regulators in archaea. The resulting knowledge will be of interest to the archaeal community and the non-coding RNA and RNA communities.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该项目研究了小的非编码RNA（sRNA）在微生物对环境胁迫的反应中的作用。在与真核生物有着密切进化关系的微生物类群--微生物中发现了数百种sRNA，但对它们的作用机制以及它们如何通过进化调节生物体的功能知之甚少。该研究将阐明sRNA在模式古细菌中的调控格局，以解决这一知识空白。对古细菌sRNA功能和调控的深入了解将使sRNA分子作为合成生物学工具和生物系统的合理设计成为可能。更广泛的影响活动将使来自不同背景的学生在STEM职业生涯的早期参与实验室研究，特别是第一代大学生，女性和代表性不足的少数民族。学生将通过FiGURE（第一代本科生研究经验）招募，这是约翰霍普金斯大学的创新研究经验和队列建设计划。这些活动还将为研究生提供导师培训。本研究旨在获得对模式古菌Haloferax volcano ii（Hv）中sRNA调节的机械理解，以应对氧化应激。具体目标是（1）表征SHOxi sRNA介导的氧化应激反应的转录后机制，并确定SHOxi如何通过调节mRNA稳定性或干扰翻译来调节推定的靶点;（2）鉴定参与氧化应激反应sRNA与其靶点相互作用的伴侣蛋白和其他因子;和（3）阐明应激应答sRNA SHOxi在氧化应激下如何被调节（特别是测试经典古细菌转录因子调节SHOxi的假设）。该研究将采用遗传和生物化学方法，定点诱变结合sRNA-mRNA相互作用建模和高通量表型分析的组合。这些结果有望揭示古细菌sRNA结合和调节其靶点的机制，鉴定新的RNA结合蛋白，并发现sRNA介导的调控网络如何与古细菌中的转录调控因子整合。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。