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.

地球的深海是数量巨大、种类繁多的生物的家园，其中包括90%的微生物，这些微生物承受的压力高达地表压力的1000倍。负责关键生命过程的蛋白质机器，如遗传密码的转录，必须调整以在这种极端和不适宜居住的条件下工作。这种适应的机制仍然是一个谜。该项目旨在了解表面和压力适应微生物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提供对地球上生命的进化甚至起源的洞察，并使食品科学行业的新生物技术进步成为可能，例如，提高加压巴氏杀菌的安全性。参与该项目的研究生和博士后学者将接受多学科研究技能培训，并有机会指导本科生和高中生。所有受训人员都将参加极端生物学和生物物理学研究协调网络。在中嗜性大肠杆菌和中等嗜压性细菌深部光杆菌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。在大肠杆菌中，高压诱导热休克反应，反之，低压诱导深度假单胞菌SS9热休克反应。这项研究将研究压力对家务管理和应激反应RNA聚合酶西格玛因子转录调节因子启动子相互作用的影响，以及西格玛因子蛋白本身的稳定性。这些反应将在深度假单胞菌SS9和一种压力适应的大肠杆菌菌株中进行表征，以进行比较分析。还将研究ToxR/ToxS转录调节系统，该系统参与了深松假单胞菌SS9转录程序的压力调节。这项研究将涉及使用新开发的在压力下的荧光显微镜技术直接观察和量化转录蛋白质的性质和过程，并将使用随机建模来更深入地了解这些过程中特定生化步骤的压力敏感性。项目成果有望为地球高压环境的进化适应提供洞察力，并可能有助于缓解食品工业中耐压微生物造成的问题。该项目由生物科学局分子和细胞生物科学部的遗传机制和分子生物物理学项目以及地球科学局海洋科学部的生物海洋学项目共同资助。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。