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

RUI：探究贝氏不动杆菌的两个不同的双精氨酸转位酶

基本信息

批准号：
1615822
负责人：
Nathan Rigel
金额：
$ 45.49万
依托单位：
Hofstra University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-15 至 2021-04-30
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1615822&HistoricalAwards=false
关键词：
RUI Probing Two Distinct Twin

项目摘要

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.

细菌具有在各种环境中定植和繁殖的惊人能力。在这些不同的环境中，最成功的细菌可以适应不断变化的疾病，从而胜过其微生物邻居。对于居住的细菌，例如贝利菌，一种关键的生存策略依赖于导出多种蛋白质曲目，从而允许营养摄取和对有毒化合物的耐药性。该项目的长期目标是阐明特定的途径如何促进A. baylyi的蛋白质出口，尤其是与该微生物在野外有助于生存有关的途径。该项目将为霍夫斯特拉大学的本科生和硕士学生提供机会，以在分子微生物学的各个领域获得丰富的动手经验。将该研究项目的一部分整合到现有课程中，将增强Hofstra学生的教育经验，超出了典型的课堂环境。此外，附近的布伦特伍德高中的学生将参加该项目，从而向他们介绍假设驱动的研究。在所有细胞生物中都发现了蛋白质出口途径，这些途径对于维持生命至关重要。双精氨酸易位酶（TAT）途径被广泛分布为在细菌，古细菌和植物囊体中发现的同源物。因此，了解TAT途径在分子水平上的功能如何在真核和原核生物中应用。 A. Baylyi提供了一个独特的系统，可以在其中研究TAT介导的蛋白质输出。实际上，A。Baylyi中的双重TAT途径的存在表明，TAT介导的蛋白质导出比当前模型所建议的更为复杂。使用A. baylyi作为研究TAT依赖性蛋白质导出的模型将提供其他模型生物（如大肠杆菌）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。为了确定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学分析。最后，使用遗传和生化方法，他们将研究从贝利曲霉细胞纯化的TAT复合物的分子结构。