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

这一行动为NSF 2022财年生物学博士后研究奖学金提供了资金，综合研究调查了基因组、环境和表型之间相互作用的生命规则。该奖学金支持研究员的研究和培训，这些研究员将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，这是生命的一个关键方面。生物体环境的变化可能会导致基因开启和关闭的方式发生巨大差异，从而使有机体能够适应和响应不同的环境刺激。对于许多生物体来说，对环境应激源的遗传反应的潜在机制是未知的。这项提议旨在确定一种名为Thermus thermophilus的模式微生物如何控制基因的开启和关闭，以响应环境中的重金属。该项目将在以本科为主的大型大学--肯纳索州立大学进行，该大学将为这位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大量机会。嗜热微生物通常被用作实验室环境中的模型系统，它们的蛋白质在生物技术和工业中有相当大的应用。尽管它们被广泛使用，但我们对许多嗜热菌模型背后的基本生物学的了解是有限的。嗜热菌HB8含有三个铁摄取调节因子(FUR)基因同源物，其基因产物的研究严重不足。FUR蛋白家族是转录调节因子，影响基因表达以响应调节金属。本研究将使用限制性内切酶保护、选择和扩增(REPSA)的组合方法来鉴定每个Thermus thermophilus HB8毛皮同源物的转录调控网络。通过REPSA实验发现的每一种蛋白质的共识DNA结合序列将通过大规模并行半导体测序和多个Em for Motif启发式软件进行鉴定。这些基序将被映射到有机体的基因组中，使用查找单个基序出现。潜在的调控基因将通过DNA结合分析和体外转录在体外进行验证，以及通过突变的嗜热菌HB8菌株的基因表达分析在体内进行验证。这项工作的结果将提供更好的理解重金属是如何通过转录调控蛋白影响极端细菌模型中的基因表达的。这项工作还应该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，该项目将支持微生物学和生物化学领域的几名本科生和研究生的指导和培训。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.