Investigating the Antifungal Mechanism of Action of Polygodial by Phenotypic Screening in Saccharomyces cerevisiae.

Purity N Kipanga,Walter Luyten,Joris Winderickx,Geert Callewaert,Petra D’Hooge,Elja Eskes,Patrick Van Dijck,Liesbeth Demuyser,Josphat Matasyoh,Johannes Vrijdag

doi:10.3390/ijms22115756

Abstract

Polygodial is a “hot” peppery-tasting sesquiterpenoid that was first described for its anti-feedant activity against African armyworms. Using the haploid deletion mutant library of Saccharomyces cerevisiae, a genome-wide mutant screen was performed to shed more light on polygodial’s antifungal mechanism of action. We identified 66 deletion strains that were hypersensitive and 47 that were highly resistant to polygodial treatment. Among the hypersensitive strains, an enrichment was found for genes required for vacuolar acidification, amino acid biosynthesis, nucleosome mobilization, the transcription mediator complex, autophagy and vesicular trafficking, while the resistant strains were enriched for genes encoding cytoskeleton-binding proteins, ribosomal proteins, mitochondrial matrix proteins, components of the heme activator protein (HAP) complex, and known regulators of the target of rapamycin complex 1 (TORC1) signaling. WE confirm that polygodial triggers a dose-dependent vacuolar alkalinization and that it increases Ca2+ influx and inhibits glucose-induced Ca2+ signaling. Moreover, we provide evidence suggesting that TORC1 signaling and its protective agent ubiquitin play a central role in polygodial resistance, suggesting that they can be targeted by polygodial either directly or via altered Ca2+ homeostasis.

Highlights

We isolated polygodial from Warburgia ugandensis Sprague subspecies ugandensis (Canellaceae) and showed its activity against Candida albicans [6]
We explored the antifungal mechanism of action (MOA) of polygodial on C. albicans using a genome-wide cerevisiae was selected as a model organism for a number of reasons, some of which are its growth-based screening of the haploid deletion strain library of S. cerevisiae [26,27]
Phenotypic Screening of Mutants Treated with Polygodial

Summary

Introduction

Apart from the anti-feedant activity against a number of pests [1,2,3], polygodial (Figure 1) has been reported to possess biological activities such as antifungal [4,5,6], antibacterial [7], anti-tumor [8], larvicidal [9], antihelminthic [10], antifouling [11], antiinflammatory [12], analgesic [13], antitrypanosomal, and antileishmanial activities [14]. Kubo intimated a more complex mechanism of polygodial’s they showed, using isolated membrane fractions of S. membrane cerevisiae, that polygodial rapidly activity [21]. They showed, using isolated fractions of S. cerevisiae, inhibited mitochondrial. They concluded that polygodial’s major target was the results in low cellular. S. We explored the antifungal MOA of polygodial on C. albicans using a genome-wide cerevisiae was selected as a model organism for a number of reasons, some of which are its growth-based screening of the haploid deletion strain library of S. cerevisiae [26,27].

Phenotypic Screening of Mutants Treated with Polygodial

Vacuolar pH Measurements in the Presence of Polygodial

Discussion

Chemical Genetic Screen

Calcium Measurements Using an Apoaequorin System

Vacuolar pH Measurements