Vazquez-Rivera Emmanuel, Rojas Brenda L, Carney Patrick R, Marrero-Valentin Jose L, Bradfield Christopher A
Molecular and Environmental Toxicology Center, McArdle Laboratory for Cancer Research, University of Wisconsin School of Medicine and Public Health, Madison, WI 53706, USA.
Toxicol Rep. 2022 Mar 17;9:513-520. doi: 10.1016/j.toxrep.2022.03.012. eCollection 2022.
Proteins, such as the Ah receptor (AHR), hold potential as sensors to detect ligands in environmental and biological samples, and may also serve as tools to regulate biosynthetic and industrial processes. The AHR is also a prototype system for the PAS superfamily that can sense and mediate adaptation to signals as diverse as light, voltage, oxygen and an array of small molecules. The yeast, has proven to be an important model to study the signal transduction of sensors like the AHR because of its ease of use, numerous available strategies for genetic manipulation, and capacity for heterologous expression. To better understand the utility of sensor proteins as components of yeast detection systems, we characterized a chimeric AHR-LexA system that drives expression from a Lex operator (LexO) driven, beta-galactosidase (β-Gal) reporter. In this report, we demonstrate that improvements in assays sensitivity and pharmacology can arise from the careful optimization of yeast growth phase and the duration of ligand exposure. We also report that the coexpression of heterotypic modifiers from mammalian cells (e.g., the ARA9 and ARA3 proteins), can improve yeast assay performance. We propose that complementing these assay improvements with previously reported yeast mutations described by others will expand the utility of the AHR for biotechnology applications.
蛋白质，如芳烃受体（AHR），有潜力作为检测环境和生物样品中配体的传感器，也可作为调节生物合成和工业过程的工具。AHR也是PAS超家族的一个原型系统，它能感知并介导对多种信号的适应，这些信号包括光、电压、氧气和一系列小分子。酵母因其易于使用、有众多可用的基因操作策略以及异源表达能力，已被证明是研究像AHR这样的传感器信号转导的重要模型。为了更好地理解传感器蛋白作为酵母检测系统组件的效用，我们对一种嵌合AHR-LexA系统进行了表征，该系统驱动来自Lex操纵子（LexO）驱动的β-半乳糖苷酶（β-Gal）报告基因的表达。在本报告中，我们证明了通过仔细优化酵母生长阶段和配体暴露持续时间，可以提高检测灵敏度和药理学效果。我们还报告说，共表达来自哺乳动物细胞的异型修饰因子（如ARA9和ARA3蛋白）可以改善酵母检测性能。我们建议，将这些检测改进与其他人先前报道的酵母突变相结合，将扩大AHR在生物技术应用中的效用。
