Antifungal activity in <i>Saccharomyces cerevisiae</i> is modulated by calcium signalling

Edlind, Thomas D.; Smith, Lamar; Henry, Karl W.; Katiyar, Säntosh K.; Nickels, Joseph T.

doi:10.1046/j.1365-2958.2002.03165.x

Cited by 112 publications

(95 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Trp was shown to mitigate the antifungal activity of ibuprofen, an anti-inflammatory agent with antifungal action, and of the calcineurin inhibitor FK506, an immunosuppressant with antifungal activity (16,18). Interestingly, we also found the calcineurin cnb1 deletion mutant to be hypersensitive to miconazole, as was previously demonstrated by Edlind et al (19). However, it is not clear how Trp can protect yeast cells against various stresses.…”

Section: Discussionsupporting

confidence: 78%

Miconazole Induces Changes in Actin Cytoskeleton prior to Reactive Oxygen Species Induction in Yeast

Thevissen

Ayscough

Aerts

et al. 2007

Journal of Biological Chemistry

View full text Add to dashboard Cite

The antifungal compound miconazole inhibits ergosterol biosynthesis and induces reactive oxygen species (ROS) in susceptible yeast species. To further uncover the mechanism of miconazole antifungal action and tolerance mechanisms, we screened the complete set of haploid Saccharomyces cerevisiae gene deletion mutants for mutants with an altered miconazole sensitivity phenotype. We identified 29 S. cerevisiae genes, which when deleted conferred at least 4-fold hypersensitivity to miconazole. Major functional groups encode proteins involved in tryptophan biosynthesis, membrane trafficking including endocytosis, regulation of actin cytoskeleton, and gene expression. With respect to the antifungal activity of miconazole, we demonstrate an antagonism with tryptophan and a synergy with a yeast endocytosis inhibitor. Because actin dynamics and induction of ROS are linked in yeast, we further focused on miconazole-mediated changes in actin cytoskeleton organization. In this respect, we demonstrate that miconazole induces changes in the actin cytoskeleton, indicative of increased filament stability, prior to ROS induction. These data provide novel mechanistic insights in the mode of action of a ROS-inducing azole.Miconazole belongs to the azole antifungals, which are currently the most widely used antimycotics. Azoles inhibit ergosterol biosynthesis, resulting in accumulation of toxic methylated sterol intermediates and, subsequently, fungal cell growth arrest (1). Recently, an additional mode of antifungal action was uncovered for miconazole. Apart from inhibition of ergosterol biosynthesis, miconazole induces reactive oxygen species (ROS) 6 in susceptible fungi, leading to fungal cell death (2, 3).The aim of the present study was to further unravel the mode of antifungal action of miconazole and tolerance mechanisms against miconazole in yeast. Information regarding drug tolerance mechanisms is invaluable for antifungal therapy: theoretically, miconazole action can be increased by using inhibitors of identified yeast tolerance mechanisms. To our knowledge, this study is the first report on yeast tolerance mechanisms against imidazoles, such as miconazole, and their mode of action using a genome-wide screening approach in yeast. In this study, we screened the haploid set of Saccharomyces cerevisiae deletion mutants in non-essential genes for both hypersensitivity and resistance to miconazole by determining the minimal inhibitory miconazole concentration (MIC) for all individual yeast knock-out mutants using 2-fold dilution series of miconazole in liquid YPD medium. In this way, no miconazole-resistant yeast mutants could be identified. However, we could identify 29 S. cerevisiae genes or open reading frames that upon deletion result in at least 4-fold hypersensitivity to miconazole and hence are involved in miconazole tolerance. The following major functional families of miconazole tolerance genes could be distinguished: (i) tryptophan biosynthesis, (ii) membrane trafficking, including endocytosis, (iii) regulation of ac...

show abstract

Section: Discussionsupporting

confidence: 78%

Miconazole Induces Changes in Actin Cytoskeleton prior to Reactive Oxygen Species Induction in Yeast

Thevissen

Ayscough

Aerts

et al. 2007

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…We verified this idea as we showed that activation of FKS2, a well established reporter gene of cell wall defects (6,17,29), in response to Calcofluor White or in a gas1 mutant was reduced upon the removal of the CDRE element present in the promoter of this gene. Interestingly, two recent papers reported the implication of calcium signaling in the tolerance of yeast to antifungal azoles (96) and in transcriptional induction of genes in response to alkaline pH (97). Altogether, these data demonstrate a major contribution of the calcium signaling in adaptation of yeast cells to various stresses.…”

Section: Discussionmentioning

confidence: 67%

Genome-wide Analysis of the Response to Cell Wall Mutations in the Yeast Saccharomyces cerevisiae

Lagorce

Hauser

Labourdette

et al. 2003

Journal of Biological Chemistry

241

280

View full text Add to dashboard Cite

Perturbations of the yeast cell wall trigger a repair mechanism that reconfigures its molecular structure to preserve cell integrity. To investigate this mechanism, we compared the global gene expression in five mutant strains, each bearing a mutation (i.e. fks1, kre6, mnn9, gas1, and knr4 mutants) that affects in a different manner the cell wall construction. Altogether, 300 responsive genes were kept based on high stringency criteria during data processing. Functional classification of these differentially expressed genes showed a substantial subset of induced genes involved in cell wall construction and an enrichment of metabolic, energy generation, and cell defense categories, whereas families of genes belonging to transcription, protein synthesis, and cellular growth were underrepresented. Clustering methods isolated a single group of ϳ80 up-regulated genes that could be considered as the stereotypical transcriptional response of the cell wall compensatory mechanism. The in silico analysis of the DNA upstream region of these co-regulated genes revealed pairwise combinations of DNA-binding sites for transcriptional factors implicated in stress and heat shock responses (Msn2/4p and Hsf1p) with Rlm1p and Swi4p, two PKC1-regulated transcription factors involved in the activation genes related to cell wall biogenesis and G 1 /S transition. Moreover, this computational analysis also uncovered the 6-bp 5 -AGCCTC-3 CDRE (calcineurindependent response element) motif in 40% of the coregulated genes. This motif was recently shown to be the DNA binding site for Crz1p, the major effector of calcineurin-regulated gene expression in yeast. Taken altogether, the data presented here lead to the conclusion that the cell wall compensatory mechanism, as triggered by cell wall mutations, integrates three major regulatory systems: namely the PKC1-SLT2 mitogen-activated protein kinase-signaling module, the "global stress" response mediated by Msn2/4p, and the Ca 2؉ /calcineurindependent pathway. The relative importance of these regulatory systems in the cell wall compensatory mechanism is discussed.Yeast and fungi are surrounded by a cell wall that is a complex structure essential for maintenance of the cell shape, prevention of lysis, and protection against harmful environmental conditions. The yeast cell wall architecture has been determined in detail over the past decade (1). It is a layered structure that is composed of ␤-1,3-and ␤-1,6-glucan (50 -60% of the cell wall dry mass), mannoproteins (40 -50%), and chitin (2%). ␤-1,3-Glucan and chitin form a fibrillar network to which mannoproteins are anchored, mostly through ␤-1,6-glucan. Some cell wall proteins, like the PIR family, can be directly linked to ␤-1,3-glucan (reviewed in Ref.2). The cell wall is not a rigid structure, since it endures all of the changes that the cell undergoes during division, morphogenesis, and differentiation.To ensure continuous integrity of the wall in accordance with its plasticity, complex mechanisms must be operating, which need to be strictly ...

show abstract

“…Relatedly, the signaling pathways that mediate phenotypic adaptation to antifungal treatment are just beginning to be understood. Recent studies have implicated the calcium-calmodulin-calcineurin (7,11,24,26) and protein kinase C (PKC)-cell integrity (T. D. Edlind, Abstr. 41st Intersci.…”

mentioning

confidence: 99%

Cyclic AMP Signaling Pathway Modulates Susceptibility of Candida Species and Saccharomyces cerevisiae to Antifungal Azoles and Other Sterol Biosynthesis Inhibitors

Jain

Akula

Edlind

2003

Antimicrob Agents Chemother

Self Cite

View full text Add to dashboard Cite

Azoles are widely used antifungals; however, their efficacy is compromised by fungistatic activity and selection of resistant strains during treatment. Recent studies demonstrated roles for the protein kinase C and calcium signaling pathways in modulating azole activity. Here we explored a role for the signaling pathway mediated by cyclic AMP (cAMP), which is synthesized by the regulated action of adenylate cyclase (encoded by CDC35 in Candida albicans and CYR1 in Saccharomyces cerevisiae) and cyclase-associated protein (encoded by CAP1 and SRV2, respectively). Relative to wild-type strains, C. albicans and S. cerevisiae strains mutated in these genes were hypersusceptible to fluconazole (>4-to >16-fold-decreased 48-h MIC), itraconazole (>8-to >64-fold), or miconazole (16-to >64-fold). Similarly, they were hypersusceptible to terbinafine and fenpropimorph (2-to >16-fold), which, like azoles, inhibit sterol biosynthesis. Addition of cAMP to the medium at least partially reversed the hypersusceptibility of Ca-cdc35 and Sc-cyr1-2 mutants. An inhibitor of mammalian adenylate cyclase, MDL-12330A, was tested in combination with azoles; a synergistic effect was observed against azole-susceptible and -resistant strains of C. albicans and five of six non-C. albicans Candida species. Analysis of cAMP levels after glucose induction in the presence and absence of MDL-12330A confirmed that it acts by inhibiting cAMP synthesis in yeast. RNA analysis suggested that a defect in azole-dependent upregulation of the multidrug transporter gene CDR1 contributes to the hypersusceptibility of the Ca-cdc35 mutant. Our results implicate cAMP signaling in the yeast azole response; compounds similar to MDL-12330A may be useful adjuvants in azole therapy.

show abstract

Antifungal activity in Saccharomyces cerevisiae is modulated by calcium signalling

Cited by 112 publications

References 47 publications

Miconazole Induces Changes in Actin Cytoskeleton prior to Reactive Oxygen Species Induction in Yeast

Miconazole Induces Changes in Actin Cytoskeleton prior to Reactive Oxygen Species Induction in Yeast

Genome-wide Analysis of the Response to Cell Wall Mutations in the Yeast Saccharomyces cerevisiae

Cyclic AMP Signaling Pathway Modulates Susceptibility of Candida Species and Saccharomyces cerevisiae to Antifungal Azoles and Other Sterol Biosynthesis Inhibitors

Contact Info

Product

Resources

About