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

LysR-转录调节因子(Lttrs)是细菌中最大的调节因子家族。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LTTRs在允许细菌对环境做出反应方面发挥了核心作用，但它们是研究最少的调节器类型之一。这个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：Lttrs如何与特定的DNA序列结合，以响应特定的控制分子，以及当它们结合它们的控制分子时，会发生什么构象变化？将开发研究LTTrs的新方法，以便在科学界推进对这些蛋白质及其潜在有益应用的理解。这两个PI的联合实验室将专注于来自土壤细菌贝利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射线结晶学。这项研究将允许开发模型来解释不同的lttrs如何发挥作用。这些研究的应用可能导致新的生物传感器和/或用于生物处理和生物修复的改良生物，这些生物可以通过调节它们的功能而产生。研究环境对教育有很高的影响。该项目将使高中生、本科生和研究生有机会在一个跨越多个学科的高度协作的环境中体验实验科学和学习先进方法。

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.