A synthetic platform for multiplex screening of cell-cell communication between soil fungi

Schiesaro, G.; Undabarrena, A.; Sorensen, M. B.; Wäneskog, M.; Jönsson, M.; Tenente, R.; Rago, D.; Deichmann, M.; Hoch-Schneider, E.; Kandasamy, V.; Özdemir, E.; Cruz-Morales, P.; Frimurer, T. M.; Clemmensen, L. K. H.; Jensen, M. K.; Jensen, E. D.

doi:10.1101/2024.10.30.620951

bioRxiv · 10.1101/2024.10.30.620951

A synthetic platform for multiplex screening of cell-cell communication between soil fungi

Schiesaro, G.·Undabarrena, A.·Sorensen, M. B.·Wäneskog, M.·Jönsson, M.·Tenente, R.·Rago, D.·Deichmann, M.·Hoch-Schneider, E.·Kandasamy, V.·Özdemir, E.·Cruz-Morales, P.·Frimurer, T. M.·Clemmensen, L. K. H.·Jensen, M. K.·Jensen, E. D.

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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 new 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 signalled 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 model system for studying fungal cell-cell communication.

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Schiesaro, G., Undabarrena, A., Sorensen, M. B., Wäneskog, M., Jönsson, M., Tenente, R., Rago, D., Deichmann, M., Hoch-Schneider, E., Kandasamy, V., Özdemir, E., Cruz-Morales, P., Frimurer, T. M., Clemmensen, L. K. H., Jensen, M. K., Jensen, E. D.. 2024-10-30. A synthetic platform for multiplex screening of cell-cell communication between soil fungi. https://doi.org/10.1101/2024.10.30.620951

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