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.<br/><br/>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.