RUI: Probing the Two Distinct Twin-Arginine Translocases of Acinetobacter baylyi

细菌有惊人的能力在不同的环境中繁衍生息。在这些不同的环境中，最成功的细菌可以适应不断变化的条件，从而胜过它们的微生物邻居。对于像baylyacinetobacter baylyi这样生活在土壤中的细菌来说，一个关键的生存策略依赖于输出多种蛋白质，从而吸收营养并抵抗有毒化合物。该项目的长期目标是阐明baylyi的特定途径如何有助于输出蛋白质，特别是因为它与帮助这种微生物在野外生存有关。该项目将为霍夫斯特拉大学的本科生和硕士生提供机会，在分子微生物学的各个领域获得重要的实践经验。将该研究项目的一部分整合到现有课程中，将增强霍夫斯特拉大学学生在典型课堂环境中获得的教育体验。此外，来自附近布伦特伍德高中的学生将参与该项目，从而向他们介绍假设驱动的研究。蛋白质输出途径存在于所有细胞生物中，这些途径对维持生命至关重要。双精氨酸转位酶（Twin-Arginine Translocase, Tat）途径广泛分布于细菌、古细菌和植物类囊体中。因此，了解Tat途径在分子水平上的功能对真核生物和原核生物都有应用价值。baylyi提供了一个独特的系统来研究tat介导的蛋白质输出。事实上，巴氏拟南芥中双Tat通路的存在表明，Tat介导的蛋白质输出比目前模型所显示的更为复杂。利用a . baylyi作为模型来研究tat依赖性蛋白质的输出将提供在大肠杆菌等其他模式生物中无法获得的见解。特别是，研究人员将研究对生长重要的表型，包括依赖于Tat的生物膜形成和细胞包膜生物发生。为了鉴定baylyi中所有Tat底物，研究人员将使用基于候选物的方法和全面的蛋白质组学分析。最后，利用遗传和生化方法，他们将研究从baylyi细胞中纯化的Tat复合物的分子结构。

Bacteria have the amazing ability to colonize and thrive in diverse environments. Within these diverse environments, the most successful bacteria can adapt to changing conditions and thereby outcompete their microbial neighbors. For soil-dwelling bacteria like Acinetobacter baylyi, one key survival strategy relies on exporting a diverse repertoire of proteins allowing nutrient uptake and resistance to toxic compounds. The long-term goal of this project is to clarify how specific pathways contribute to exporting proteins of A. baylyi, particularly as it relates to aiding survival of this microbe in the wild. This project will provide opportunities for undergraduate and masters students at Hofstra University to gain significant hands-on experience in various areas of molecular microbiology. Integrating part of this research project into existing courses will enhance the educational experience of Hofstra students beyond that obtained in a typical classroom setting. Additionally, students from nearby Brentwood High School will participate in the project thereby introducing them to hypothesis-driven research.Protein export pathways are found in all cellular organisms, and these pathways are critical for sustaining life. The Twin-Arginine Translocase (Tat) pathway is widely distributed as homologs of it are found in bacteria, archaea, and plant thylakoids. Therefore, understanding how the Tat pathway functions at a molecular level has applications for both eukaryotic and prokaryotic organisms. A. baylyi provides a unique system in which to study Tat-mediated protein export. Indeed, the existence of dual Tat pathways in A. baylyi indicates that Tat-mediated protein export is more complicated than current models suggest. Using A. baylyi as a model to study Tat-dependent protein export will provide insights not available in other model organisms like Escherichia coli. In particular, the researchers will investigate phenotypes important for growth including biofilm formation and cell envelope biogenesis that are dependent on Tat. To identify all Tat substrates in A. baylyi, the investigators will use both a candidate-based approach and a comprehensive proteomic analysis. Lastly, using both genetic and biochemical approaches, they will investigate the molecular architecture of Tat complexes purified from A. baylyi cells.