High-Content, Image-Based Screening for Drug Targets in Yeast

Ohnuki, Shinsuke; Oka, Satomi; Nogami, Satoru; Ohya, Yoshikazu

doi:10.1371/journal.pone.0010177

Cited by 47 publications

(61 citation statements)

References 45 publications

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“…To assess the morphological similarity between the cells treated with poacic acid and nonessential deletion mutants or cells treated with other cell wall-affecting drugs, their morphological profiles were compared as described previously (36). To identify functional gene clusters, the most significant similar mutants (43 genes, P < 0.01 after Bonferroni correction, t test) were selected as a query for GO term analysis (GO term finder in the Saccharomyces genome database).…”

Section: Methodsmentioning

confidence: 99%

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Plant-derived antifungal agent poacic acid targets β-1,3-glucan

Piotrowski

Okada

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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136

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A rise in resistance to current antifungals necessitates strategies to identify alternative sources of effective fungicides. We report the discovery of poacic acid, a potent antifungal compound found in lignocellulosic hydrolysates of grasses. Chemical genomics using Saccharomyces cerevisiae showed that loss of cell wall synthesis and maintenance genes conferred increased sensitivity to poacic acid. Morphological analysis revealed that cells treated with poacic acid behaved similarly to cells treated with other cell wall-targeting drugs and mutants with deletions in genes involved in processes related to cell wall biogenesis. Poacic acid causes rapid cell lysis and is synergistic with caspofungin and fluconazole. The cellular target was identified; poacic acid localized to the cell wall and inhibited β-1,3-glucan synthesis in vivo and in vitro, apparently by directly binding β-1,3-glucan. Through its activity on the glucan layer, poacic acid inhibits growth of the fungi Sclerotinia sclerotiorum and Alternaria solani as well as the oomycete Phytophthora sojae. A single application of poacic acid to leaves infected with the broad-range fungal pathogen S. sclerotiorum substantially reduced lesion development. The discovery of poacic acid as a natural antifungal agent targeting β-1,3-glucan highlights the potential side use of products generated in the processing of renewable biomass toward biofuels as a source of valuable bioactive compounds and further clarifies the nature and mechanism of fermentation inhibitors found in lignocellulosic hydrolysates.

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Section: Methodsmentioning

confidence: 99%

“…We investigated the morphological changes induced by poacic acid using high-dimensional microscopy (36,37). Recently, two morphological features (a wide neck and morphological heterogeneity) were reported as common phenotypes in cells treated with agents known to affect the cell wall (38).…”

Section: Morphological Analysis Revealed That Poacic Acid Affects Thementioning

confidence: 99%

Plant-derived antifungal agent poacic acid targets β-1,3-glucan

Piotrowski

Okada

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

136

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“…The Box-Cox transformation method uses maximum likelihood to determine which among a family of power transformations produces a phenotypic distribution that approximates normality best [60]. Previous studies using CalMorph data have used this transformation [47], [61]–[63]. If a phenotype did not show an approximately normal distribution on a qq-plot after this procedure, it was excluded (the phenotypes used in the analysis are shown in Table S6).…”

Section: Methodsmentioning

confidence: 99%

Histone Variant HTZ1 Shows Extensive Epistasis with, but Does Not Increase Robustness to, New Mutations

et al. 2013

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Biological systems produce phenotypes that appear to be robust to perturbation by mutations and environmental variation. Prior studies identified genes that, when impaired, reveal previously cryptic genetic variation. This result is typically interpreted as evidence that the disrupted gene normally increases robustness to mutations, as such robustness would allow cryptic variants to accumulate. However, revelation of cryptic genetic variation is not necessarily evidence that a mutationally robust state has been made less robust. Demonstrating a difference in robustness requires comparing the ability of each state (with the gene perturbed or intact) to suppress the effects of new mutations. Previous studies used strains in which the existing genetic variation had been filtered by selection. Here, we use mutation accumulation (MA) lines that have experienced minimal selection, to test the ability of histone H2A.Z (HTZ1) to increase robustness to mutations in the yeast Saccharomyces cerevisiae. HTZ1, a regulator of chromatin structure and gene expression, represents a class of genes implicated in mutational robustness. It had previously been shown to increase robustness of yeast cell morphology to fluctuations in the external or internal microenvironment. We measured morphological variation within and among 79 MA lines with and without HTZ1. Analysis of within-line variation confirms that HTZ1 increases microenvironmental robustness. Analysis of between-line variation shows the morphological effects of eliminating HTZ1 to be highly dependent on the line, which implies that HTZ1 interacts with mutations that have accumulated in the lines. However, lines without HTZ1 are, as a group, not more phenotypically diverse than lines with HTZ1 present. The presence of HTZ1, therefore, does not confer greater robustness to mutations than its absence. Our results provide experimental evidence that revelation of cryptic genetic variation cannot be assumed to be caused by loss of robustness, and therefore force reevaluation of prior claims based on that assumption.

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“…High-content microscopy has also been used as a tool to decipher MOA of bioactive compounds [20–22]. By monitoring a myriad of parameters from a cell exposed to a chemical agent (target expression, localization, cell morphology, organelle function, and cytotoxicity/cell health to name few), understanding that agent’s MOA can become more straightforward [22,23]. Joseph and colleagues use of a high-content assay to profile the drug-induced mechanisms of apoptosis illustrates the mutli-parametric paradigm [24].…”

Section: High-content and The Pursuit Of Phenotypic Screeningmentioning

confidence: 99%

Innovation in academic chemical screening: filling the gaps in chemical biology

Hasson

Inglese

2013

Current Opinion in Chemical Biology

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Academic screening centers across the world have endeavored to discover small molecules that can modulate biological systems. To increase the reach of functional-genomic and chemical screening programs, universities, research institutes, and governments have followed their industrial counterparts in adopting high-throughput paradigms. As academic screening efforts have steadily grown in scope and complexity, so have the ideas of what is possible with the union of technology and biology. This review addresses the recent conceptual and technological innovation that has been propelling academic screening into its own unique niche. In particular, high-content and whole-organism screening is changing how academics search for novel bioactive compounds. Importantly, we recognize examples of successful chemical probe development that have punctuated the changing technology landscape.

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High-Content, Image-Based Screening for Drug Targets in Yeast

Cited by 47 publications

References 45 publications

Plant-derived antifungal agent poacic acid targets β-1,3-glucan

Plant-derived antifungal agent poacic acid targets β-1,3-glucan

Histone Variant HTZ1 Shows Extensive Epistasis with, but Does Not Increase Robustness to, New Mutations

Innovation in academic chemical screening: filling the gaps in chemical biology

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