Structure and Function of BenM and CatM, Bacterial LysR-type Transcriptional Regulators

LysR-转录调节子（LTTR）是细菌中最大的调节子家族。 它们控制多种代谢功能，包括氨基酸生物合成、毒力、硝酸盐固定、复制和许多其他主要过程。 尽管它们在允许细菌对其环境做出反应方面发挥着核心作用，但LTTR是研究最少的调节剂之一。 该项目的目标是扩大我们对这些蛋白质如何在原子水平上发挥作用的理解。 将解决的问题包括：如何做LTTRs结合到特定的DNA序列，以响应特定的控制分子和什么样的构象变化发生时，他们结合他们的控制分子？ 将开发研究LTTR的新方法，以便在科学界进一步了解这些蛋白质及其潜在的有益应用。 这两个PI的联合实验室将专注于来自土壤细菌Baylyi不动杆菌的两种蛋白质（BenM和CatM）。将使用广泛的方法，包括突变体筛选（遗传学），DNA足迹（分子生物学）和生化方法，如色氨酸荧光，荧光偏振和X射线晶体学。这项研究将允许开发模型来解释不同的LTTR如何发挥作用。 这些研究的应用可能会导致新的生物传感器和/或改进的生物处理和生物修复，可以来自能够调节其功能。 研究环境具有很高的教育影响力。 该项目将允许高中，本科和研究生有机会体验实验科学，并在跨多个学科的高度协作环境中学习先进的方法。

LysR-Transcriptional Regulators (LTTRs) are the largest family of regulators in bacteria. They control diverse metabolic functions including amino acid biosynthesis, virulence, nitrate fixation, replication, and many other major processes. Despite their central role in allowing bacteria to respond to their environment, LTTRs are among the least studied types of regulators. The goal of this project is to expand our understanding of how these proteins function at an atomic level. Questions that will be addressed include: how do LTTRs bind to specific sequences of DNA in response to specific control molecules and what conformational changes take place when they bind their control molecules? New approaches to study LTTRs will be developed so that understanding these proteins and their potential beneficial applications can be advanced in the scientific community. The combined labs of the two PIs will focus on two proteins (BenM and CatM) from a soil bacterium, Acinetobacter baylyi. A wide range of methods will be used including mutant screens (genetics), DNA footprinting (molecular biology), and biochemical methods like tryptophan fluorescence, fluorescence polarization, and X-ray crystallography. The research will allow models to be developed to explain how diverse LTTRs function. Application of these studies may lead to novel biosensors and/or improved organisms for bioprocessing and bioremediation that can come from being able to modulate their functions. The research environment has a high educational impact. The project will allow high school, undergraduate and graduate students to have opportunities to experience experimental science and learn advanced methods in a highly collaborative environment that crosses multiple disciplines.