A yeast mating platform for multiplex screening of fungal GPCR-ligand interactions.

Abstract

Fungi are essential members across ecosystems, yet phytopathogenic fungi pose an increasing risk to crop yields. Despite their ecologic importance, cell-cell communication in fungi is underexplored, partly due to the lack of high-throughput techniques. Here, we developed a Yeast Mating Platform (YeMaP) to investigate the interaction between fungal G protein-coupled receptors (GPCRs) and pheromone peptides. We used YeMaP for high-throughput screening of 8,000 pheromone sequences and identified peptides with improved agonism or antagonism action. We found that these peptides can be applied in a native fungal system such as the plant pathogen <i>Fusarium oxysporum</i>, to control hyphal chemotropism and reduce plant root penetration. Additionally, we utilized YeMaP in a one-pot assay to investigate how abiotic factors influence the communication of multiple pheromone-GPCR combinations and found that the cell-cell communication mediated by the GPCR Ste2 from <i>F. oxysporum</i> signaled robustly across different abiotic factors, while other fungal GPCR-pheromone interactions were more sensitive to changes. Taken together, YeMaP accelerates the identification of fungal GPCR-peptide interactions by enabling one-pot assays, and serves as a model system for studying fungal cell-cell communication.