Mediator Subunit Med15 Regulates Cell Morphology and Mating in Candida lusitaniae

Abstract

Candida lusitaniae is an emerging opportunistic pathogenic yeast capable of shifting from yeast to pseudohyphae form, and it is one of the few Candida species with the ability to reproduce sexually. In this study, we showed that a dpp3Δ mutant, inactivated for a putative pyrophosphatase, is impaired in cell separation, pseudohyphal growth and mating. The defective phenotypes were not restored after the reconstruction of a wild-type DPP3 locus, reinforcing the hypothesis of the presence of an additional mutation that we suspected in our previous study. Genetic crosses and genome sequencing identified an additional mutation in MED15, encoding a subunit of the mediator complex that functions as a general transcriptional co-activator in Eukaryotes. We confirmed that inactivation of MED15 was responsible for the defective phenotypes by rescuing the dpp3Δ mutant with a wild-type copy of MED15 and constructing a med15Δ knockout mutant that mimics the phenotypes of dpp3Δ in vitro. Proteomic analyses revealed the biological processes under the control of Med15 and involved in hyphal growth, cell separation and mating. This is the first description of the functions of MED15 in the regulation of hyphal growth, cell separation and mating, and the pathways involved in C. lusitaniae.

The role of the Mediator complex in fungal pathogenesis and response to antifungal agents

“…Proteomic analysis revealed that this mutation led to the up-regulation of genes involved in stress responses and utilization of alternative carbon sources. Indirect data from this study also suggest that Med15 may contribute to, but is not essential for, wild-type levels of virulence [ 28 ]. Screening a mutant library of the filamentous fungal pathogen Aspergillus fumigatus revealed that down-regulation of a gene, AfMED15 , encoding a protein with a fungal Med15 domain, affected a wide range of phenotypes, including conidiation and virulence in the Galleria mellonella wax moth infection model.…”