Engineering a Model Cell for Rational Tuning of GPCR Signaling

• Published in: Cell 2019-04, Vol.177 (3), p.782-796.e27
• Main Authors: Shaw, William M., Yamauchi, Hitoshi, Mead, Jack, Gowers, Glen-Oliver F., Bell, David J., Öling, David, Larsson, Niklas, Wigglesworth, Mark, Ladds, Graham, Ellis, Tom
• Format: Article
• Language: English
• Subjects: biosensor; cell signaling; cell-to-cell communication; G protein-coupled receptor; Gene Expression - drug effects; Genetic Engineering; genome engineering; Humans; Pheromones - pharmacology; Receptors, G-Protein-Coupled - metabolism; Saccharomyces cerevisiae; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Signal Transduction - drug effects; synthetic biology; Transcription Factors - genetics; Transcription Factors - metabolism
• ISSN: 0092-8674; 1097-4172
• DOI: 10.1016/j.cell.2019.02.023

G protein-coupled receptor (GPCR) signaling is the primary method eukaryotes use to respond to specific cues in their environment. However, the relationship between stimulus and response for each GPCR is difficult to predict due to diversity in natural signal transduction architecture and expression. Using genome engineering in yeast, we constructed an insulated, modular GPCR signal transduction system to study how the response to stimuli can be predictably tuned using synthetic tools. We delineated the contributions of a minimal set of key components via computational and experimental refactoring, identifying simple design principles for rationally tuning the dose response. Using five different GPCRs, we demonstrate how this enables cells and consortia to be engineered to respond to desired concentrations of peptides, metabolites, and hormones relevant to human health. This work enables rational tuning of cell sensing while providing a framework to guide reprogramming of GPCR-based signaling in other systems. [Display omitted] •Genomic refactoring of a minimized GPCR signaling pathway in S. cerevisiae•Altering expression levels of key signaling components provides pathway tunability•Engineered microbial consortia provide desired dose-response characteristics•Development of fine-tuned sensors for detecting molecules relevant to human health Engineering GPCR-based signaling in yeast allows exploration of signaling properties and creation of synthetic responses to a range of ligands.