Collaborative Research: Transcriptional Adaptation and Response to Pressure

合作研究：转录适应和对压力的反应

• 批准号: 2019455
• 负责人: Douglas Bartlett
• 金额: $ 46.27万
• 依托单位: University of California-San Diego Scripps Inst of Oceanography
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2019455&HistoricalAwards=false
• 关键词: Collaborative; Research; Transcriptional; Adaptation; Response

The Earth’s deep oceans are home to an enormous amount and diversity of living creatures, including ~90% of the microbes, which can experience pressures as high as 1000 times that on the surface. The protein machines responsible for critical life processes, such as transcription of the genetic code, must be adapted to function under such extreme and inhospitable conditions. The mechanisms of this adaptation remain a mystery. This project is aimed at understanding molecular mechanisms underlying the response and adaptation to high pressure of the transcription machinery of surface- and pressure-adapted microorganisms. Better knowledge of these mechanisms will provide insights into the evolution and even the origins of life on Earth, and enable new biotechnological advances in the food science industry, e.g., to improve the safety of pressure pasteurization. Graduate students and postdoctoral scholars involved in the project will be trained in multidisciplinary research skills, and have the opportunity to mentor undergraduates and high school students. All trainees will participate in a Research Coordination Network on Extreme Biology and Biophysics.In the mesophile, Escherichia coli, and in the moderate piezophile, Photobacterium profundum SS9, the molecular basis of transcriptional response and adaptation to pressure involves the heat shock response system. High pressure induces a heat shock response in E. coli, and conversely, low pressure induces a heat shock response P. profundum SS9. The research will investigate pressure effects on promoter interactions of the transcriptional regulators of housekeeping and stress response RNA polymerase sigma factors, as well as the stability of the sigma factor proteins themselves. These responses will be characterized in P. profundum SS9 and a pressure-adapted strain of E. coli for comparative analysis. The ToxR/ToxS transcriptional regulator system, which is implicated in pressure regulation of the transcriptional program in P. profundum SS9, will also be examined. The research will involve direct observation and quantification of transcriptional protein properties and processes, using newly developed fluorescence microscopy techniques under pressure, and will employ stochastic modeling for deeper insights into the pressure sensitivity of specific biochemical steps in these processes. The project outcomes are expected to provide insights into evolutionary adaptation to high pressure environments on Earth and may help to mitigate problems caused by pressure resistant microorganisms in the food industry.This project is jointly funded by the Genetic Mechanisms and Molecular Biophysics programs of the Molecular and Cellular Biosciences Division in the Biological Sciences Directorate, and the Biological Oceanography program of the Ocean Sciences Division in the Geosciences Directorate.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.

地球深海是数量巨大、种类繁多的生物的家园，其中包括约 90% 的微生物，它们承受的压力高达表面的 1000 倍。负责转录等关键生命过程的蛋白质机器。这种适应机制仍然是一个谜，该项目旨在了解表面和转录机制对高压的反应和适应的分子机制。更好地了解这些机制将有助于深入了解地球上生命的进化甚至起源，并促进食品科学行业的新生物技术进步，例如提高压力巴氏灭菌的安全性。参与该项目的学者将接受多学科研究技能的培训，并有机会指导本科生和高中生。所有学员都将参与极端生物学和生物物理学的研究协调网络。嗜温菌、大肠杆菌和中度嗜压菌、深光杆菌 SS9 中，转录反应和压力适应的分子基础涉及热休克反应系统。高压会在大肠杆菌中诱导热休克反应，相反，低压会诱导热休克反应。热休克反应 P. profundum SS9 该研究将研究压力对管家转录调节因子和应激反应 RNA 聚合酶 sigma 因子的启动子相互作用的影响。这些反应将在 P. profundum SS9 和压力适应菌株 E. coli 中进行比较分析，该系统与转录的压力调节有关。 P. profundum SS9 的计划也将受到审查，该研究将涉及在压力下使用新开发的荧光显微镜技术直接观察和量化转录蛋白特性和过程，并将采用随机方法。建模以更深入地了解这些过程中特定生化步骤的压力敏感性，该项目的成果预计将提供对地球高压环境的进化适应的见解，并可能有助于减轻食品工业中耐压微生物引起的问题。该项目由生物科学局分子和细胞生物科学处遗传机制和分子生物物理学项目和地球科学局海洋科学处生物海洋学项目共同资助。授予 NSF 的法定使命，并通过评估反映使用基金会的智力优点和更广泛的影响审查标准，被认为值得支持。