Conservation of Cdc14 phosphatase specificity in plant fungal pathogens: implications for antifungal development

Andrew G Demarco,Mark C Hall,Amanda L Pendleton,Peipei Zhu,Monessha Nambiar,Stefan Paula,Jean Chmielewski,Kedric L Milholland,Antonella Pepe,Jennifer H Wisecaver,Daniel T Wesenberg,John J Whitney,W Andy Tao

doi:10.1038/s41598-020-68921-3

Abstract

Cdc14 protein phosphatases play an important role in plant infection by several fungal pathogens. This and other properties of Cdc14 enzymes make them an intriguing target for development of new antifungal crop treatments. Active site architecture and substrate specificity of Cdc14 from the model fungus Saccharomyces cerevisiae (ScCdc14) are well-defined and unique among characterized phosphatases. Cdc14 appears absent from some model plants. However, the extent of conservation of Cdc14 sequence, structure, and specificity in fungal plant pathogens is unknown. We addressed this by performing a comprehensive phylogenetic analysis of the Cdc14 family and comparing the conservation of active site structure and specificity among a sampling of plant pathogen Cdc14 homologs. We show that Cdc14 was lost in the common ancestor of angiosperm plants but is ubiquitous in ascomycete and basidiomycete fungi. The unique substrate specificity of ScCdc14 was invariant in homologs from eight diverse species of dikarya, suggesting it is conserved across the lineage. A synthetic substrate mimetic inhibited diverse fungal Cdc14 homologs with similar low µM Ki values, but had little effect on related phosphatases. Our results justify future exploration of Cdc14 as a broad spectrum antifungal target for plant protection.

Highlights

Cdc[14] protein phosphatases play an important role in plant infection by several fungal pathogens
A similar phenotype coupled with defective conidiation and reduced virulence was observed upon CDC14 deletion in the entomopathogenic fungus Beauveria bassiana[14], and CDC14 deletion in the opportunistic human pathogen Candida albicans resulted in cytokinesis defects and reduced hyphal growth required for infection[15]
CDC14 is broadly conserved in plant fungal pathogens but absent from angiosperms

Summary

Introduction

Cdc[14] protein phosphatases play an important role in plant infection by several fungal pathogens. This and other properties of Cdc[14] enzymes make them an intriguing target for development of new antifungal crop treatments. The extent of conservation of Cdc[14] sequence, structure, and specificity in fungal plant pathogens is unknown. The Cdc[14] phosphatases, known best for roles in counteracting cyclin-dependent kinase activity during mitosis in model fungi like Saccharomyces cerevisiae and Schizosaccharomyces pombe[9,10] may be an attractive novel target for development of broad-acting antifungal agents. Treatments targeting Cdc[14] might be predicted to have little adverse effect on plants or on animals consuming treated plant products

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Conclusion