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 Fusarium oxysporum , 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 F. oxysporum 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.
Article Details
Journal Info
Proceedings of the National Academy of Sciences
National Academy of Sciences
Authors (16)
Giovanni Schiesaro
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
Melani Mariscal
Departamento de Genética, Campus de Excelencia Internacional Agroalimentario, Universidad de Córdoba
Mathias Jönsson
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
Ricardo Tenente
Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen
Mathies Brinks Sørensen
Department of Applied Mathematics and Computer Science, Technical University of Denmark
Marcus Wäneskog
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
María Victoria Aguilar-Pontes
Departamento de Genética, Campus de Excelencia Internacional Agroalimentario, Universidad de Córdoba
Agustina Undabarrena
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
Marcus Deichmann
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
Emma E. Hoch-Schneider
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
Viji Kandasamy
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
Thomas M. Frimurer
Antonio Di Pietro
Departamento de Genética, Campus de Excelencia Internacional Agroalimentario, Universidad de Córdoba
Line Katrine Harder Clemmensen
Department of Applied Mathematics and Computer Science, Technical University of Denmark
Michael Krogh Jensen
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
Emil Damgaard Jensen
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark