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

细菌具有在不同环境中定居和繁衍的惊人能力。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，从而超过它们的微生物邻居。对于像贝利不动杆菌这样居住在土壤中的细菌来说，一个关键的生存策略依赖于输出一系列不同的蛋白质，使其能够吸收营养并抵抗有毒化合物。该项目的长期目标是阐明特定的途径如何有助于输出贝利沙门氏菌的蛋白质，特别是当它与帮助这种微生物在野外生存有关时。这个项目将为霍夫斯特拉大学的本科生和硕士学生提供机会，在分子微生物学的各个领域获得重要的实践经验。将这一研究项目的一部分整合到现有的课程中，将增强霍夫斯特拉学生的教育体验，而不是在典型的课堂环境中获得的体验。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体中，这些途径对维持生命至关重要。双精氨酸转位酶(Tat)途径广泛分布于细菌、古菌和植物类囊体中。因此，在分子水平上了解TAT途径的功能对真核生物和原核生物都有应用。A.baylyi为研究TAT介导的蛋白质输出提供了一个独特的系统。事实上，在黄曲霉中存在两条TAT通路，这表明TAT介导的蛋白质输出比目前模型所暗示的要复杂得多。使用A.baylyi作为研究TAT依赖的蛋白质输出的模型，将提供其他模式生物(如大肠杆菌)所没有的见解。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膜生物发生。为了鉴定A.baylyi中的所有TAT底物，研究人员将同时使用基于候选的方法和全面的蛋白质组分析。最后，他们将使用遗传学和生物化学方法，研究从A.baylyi细胞中纯化的TAT复合体的分子结构。

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.