A yeast mating platform for multiplex screening of fungal GPCR–ligand interactions
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.
Authors
- Thomas M. Frimurer (ORCID: https://orcid.org/0000-0002-9682-8245)
- Line Katrine Harder Clemmensen (ORCID: https://orcid.org/0000-0001-5527-5798)
- Michael K. Jensen (ORCID: https://orcid.org/0000-0001-7574-4707)
- Emma Elise Hoch-Schneider (ORCID: https://orcid.org/0000-0003-3619-4647)
- Melani Mariscal (ORCID: https://orcid.org/0000-0001-9179-1001)
- María Victoria Aguilar Pontes (ORCID: https://orcid.org/0000-0002-3174-8834)
- Emil D. Jensen (ORCID: https://orcid.org/0000-0002-8280-0946)
- Mathias Jönsson (ORCID: https://orcid.org/0000-0003-1168-5589)
- Antonio Di Pietro (ORCID: https://orcid.org/0000-0001-5930-5763)
- Marcus Deichmann (ORCID: https://orcid.org/0000-0003-3540-0576)
- Marcus Wäneskog (ORCID: https://orcid.org/0000-0003-2480-5631)
- Vijayalakshmi Kandasamy (ORCID: https://orcid.org/0000-0001-7211-8233)
- Giovanni Schiesaro (ORCID: https://orcid.org/0000-0002-6309-4337)
- Agustina Undabarrena (ORCID: https://orcid.org/0000-0001-9273-0665)
- Ricardo Tenente (ORCID: https://orcid.org/0000-0002-7970-278X)
- Mathies Brinks Sørensen (ORCID: https://orcid.org/0000-0002-4215-0512)
Institutions
- Novo Nordisk Foundation (DK)
- University of Córdoba (ES)
- Technical University of Denmark (DK)
Publication Details
- Journal
- Proceedings of the National Academy of Sciences
- Published
- 2025-10-24
- DOI
- https://doi.org/10.1073/pnas.2521198122
- Primary Topic
- Fungal Biology and Applications
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Villum Fonden
- Wenner-Gren Stiftelserna
- Novo Nordisk
- Ministerio de Ciencia e Innovación
- Novo Nordisk Fonden
- Agencia Estatal de Investigación