RUI: Examination of the Genetic Mechanisms Used to Control the Expression of Salinity-regulated Proteins in the Archaeon Haloferax volcanii

该项目旨在采用广泛的蛋白质组学、遗传学和生物信息学技术，用于进一步表征这些基因及其相应功能的研究，这些基因是适应模式盐古菌——火山盐古菌（Haloferax volcanii）变化盐度所必需的。蛋白质组学分析导致在Hfx中发现了一组盐度调节蛋白。目前正在等待对其调控机制的详细描述。在鉴定的蛋白质中，有一种类似细菌的转录激活因子，在转录水平上受NaCl浓度的调节。编码该蛋白的基因是细菌pspA基因的同源物，该基因在环境应激反应中受到调节。鉴于pspA在感知各种膜应激源中所起的不可或缺的作用，我们假设pspA可能在Hfx的低盐和/或高盐适应中发挥重要作用。volcanii。计划中的研究将解决以下问题：1)编码盐度调节蛋白的基因是否具有共同的调节机制？2) Hfx。火山PspA作为转录激活剂发挥作用，其方式类似于在细菌中看到的。3) Hfx。火山菌在低盐胁迫下诱导的基因/蛋白质与在高盐诱导条件下的相似。在这项研究中获得的信息将对Hfx如何。在其自然环境中，火山能够维持生长并在广泛的盐度范围内保持活力。此外，通过提供古细菌中应激调节PspA活性的第一个实验证据，可以阐明其与存在于细菌和真核生物领域的同源物的功能关系。这项研究的广泛影响将主要涉及到一些本科生的参与和培训，因为这项研究是在莱德大学进行的，这是一所小型的文理学院，没有理科研究生课程。

This project aims to employ a broad variety of proteomic, genetic, and bioinformatic techniques for use in studies that will further characterize those genes, and their corresponding functions, necessary for adaptation to variable salinities in the model haloarchaeon, Haloferax volcanii. Proteomic analyses have led to the discovery of a set of salinity-regulated proteins in Hfx. volcanii that now await detailed characterization of the mechanisms involved in their regulation. Among the proteins identified were a bacterial-like transcriptional activator that is regulated by NaCl concentrations at the level of transcription. The gene encoding this protein is a homolog of the bacterial pspA gene that is regulated in response to environmental stress. Given its integral role in sensing a variety of membrane stressors, it is hypothesized that pspA may play an important role in hypo- and/or hypersaline adaptation in Hfx. volcanii. Planned research will address the following questions: 1) do genes encoding salinity-regulated proteins share a common mechanisms of regulation?; 2) does Hfx. volcanii PspA function as a transcriptional activator and in a manner akin to that seen in bacteria?; 3) does Hfx. volcanii induce similar types of genes/proteins in response to low-salt stress as those seen under high-salt inducing conditions? The information obtained in this research will shed critical insight into how Hfx. volcanii is able to sustain growth and remain viable over a wide range of salinities in its natural environment. Furthermore, by providing the first experimental evidence for stress-regulated PspA activity in Archaea, its functional relationship to homologs present in bacterial and eukaryotic domains of life can be elucidated. The broader impacts of this study will chiefly involve the participation and training of a number of undergraduate research students, as this research takes place at Rider University, a small, liberal arts school with no graduate programs in the sciences.