NSF Postdoctoral Fellowship in Biology FY 2021: Regulation and response to environmental metals in thermophiles

该行动资助了2022财年的NSF生物学博士后研究奖学金，即调查基因组，环境和表型之间相互作用的生命规则的综合研究。该研究金支持研究员的研究和培训，以创新的方式为生活规则领域做出贡献。基因是如何开启和关闭的是生命的一个重要方面。生物体环境的变化会导致基因开启和关闭方式的巨大差异，从而使生物体能够适应和响应不同的环境刺激。对于许多生物体来说，对环境压力的遗传反应的机制是未知的。该提案旨在确定一种名为嗜热栖热菌的模式微生物如何控制基因如何响应环境中的重金属而打开和关闭。该项目将在主要是本科生的大型大学肯尼索州立大学进行，该大学将为研究员提供开展研究的资源和培训，以及教授和指导下一代科学家的充分机会。嗜热生物通常被用作实验室环境中的模型系统，并且它们的蛋白质在生物技术和工业中具有相当大的应用。尽管它们被广泛使用，但我们对许多模式嗜热菌背后的基本生物学的理解是有限的。嗜热栖热菌HB8含有三个铁摄取调节因子（FUR）基因同源物，其基因产物严重不足。FUR蛋白家族是转录调节因子，其响应于调节金属而影响基因表达。本研究将确定每个嗜热栖热菌HB8 FUR同源物的转录调控网络，使用组合的方法，限制性内切酶保护，选择和扩增（REPSA）。通过REPSA实验发现的每种蛋白质的共有DNA结合序列将通过大规模并行半导体测序和Multiple Em for Motif Elicitation软件进行鉴定。这些基序将使用Find Individual Motif Occurrences映射到生物体的基因组。将通过DNA结合试验和体外转录在体外验证潜在调控基因，以及通过突变嗜热栖热菌HB8菌株中的基因表达分析在体内验证潜在调控基因。这项工作的结果将提供一个更好地了解重金属如何影响基因表达，通过转录调控蛋白在一个模型极端微生物。这项工作还将产生广泛适用于转录因子研究的新方法，并将与科学界分享。最后，该项目将支持指导和培训微生物学和生物化学领域的几个本科生和研究生。该奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。

This action funds an NSF Postdoctoral Research Fellowship in Biology for FY 2022, Integrative Research Investigating the Rules of Life Governing Interactions Between Genomes, Environment and Phenotypes. The fellowship supports research and training of the fellow that will contribute to the area of Rules of Life in innovative ways. How genes are turned on and off is a critical aspect of life. Changes in an organism’s environment can lead to dramatic differences in how genes are turned on and off, allowing the organism to adapt and respond to different environmental stimuli. For many organisms, the mechanisms underlying a genetic response to an environmental stressor are unknown. This proposal aims to identify how a model microorganism called Thermus thermophilus control how genes are turned on and off in response to heavy metals in their environment. This project will be conducted at the large, primarily undergraduate university, Kennesaw State University, which will provide the fellow with resources and training to conduct research, as well as ample opportunity to teach and mentor the next generation of scientists. Thermophilic organisms are commonly used as model systems in a laboratory setting, and their proteins have considerable application in biotechnology and industry. Despite their widespread use, our understanding of the fundamental biology behind many model thermophiles is limited. Thermus thermophilus HB8 contains three Ferric Uptake Regulator (FUR) gene homologs, whose gene products are severely understudied. The FUR family of proteins are transcriptional regulators that influence gene expression in response to regulatory metals. This research will identify the transcription regulatory networks for each Thermus thermophilus HB8 FUR homolog using the combinatorial approach, Restriction Endonuclease Protection, Selection, and Amplification (REPSA). Consensus DNA-binding sequences for each protein discovered experimentally through REPSA will be identified by massively parallel semiconductor sequencing and Multiple Em for Motif Elicitation software. These motifs will be mapped to the organism’s genome using Find Individual Motif Occurrences. Potentially regulated genes will be validated in vitro through DNA-binding assays and in vitro transcription, as well as in vivo through gene expression analysis in mutant Thermus thermophilus HB8 strains. Results of this work will provide a better understanding of how heavy metals influence gene expression through transcription regulatory proteins in a model extremophile. This work should also result in novel methodologies, widely applicable to transcription factor studies, that will be shared with the scientific community. Finally, this project will support the mentorship and training of several undergraduate and graduate students in the fields of microbiology and biochemistry.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.