Atomic force microscopic study of the effects of ethanol on yeast cell surface morphology

Canetta, Elisabetta; Adya, Ashok K.; Walker, Graeme M.

doi:10.1111/j.1574-6968.2005.00089.x

Cited by 58 publications

(45 citation statements)

References 29 publications

(58 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…It is also worth noting that despite a higher elasticity, the cell size and cell shape of ethanol-treated yeasts were similar to those of untreated yeast cells, suggesting that these parameters are likely dependent on cell wall composition. Our results thus differ from the previous work of Canetta et al (23), who actually reported shrinkage of cells after incubation for more than 1 h with ethanol at a 10% or higher concentration. That we did not observe such a shrinkage in our work can most likely be ascribed to the difference in the methods of immobilizing cells for AFM analysis, as our method kept the cells alive, whereas yeast cells were fixed and dried on a glass slide in the work of Canetta et al (23).…”

Section: Discussioncontrasting

confidence: 57%

“…It is therefore the best tool for visualizing and quantifying effects of ethanol stress at the single-cell level. This question was previously approached by Canetta et al (23). However, they used an air-drying immobilization technique to fix ethanol-treated yeast cells on a glass slide.…”

mentioning

confidence: 99%

“…Thus, the immobilization technique has to be "neutral" to avoid interfering with the biological condition under study. As these conditions were apparently not respected in the work of Canetta et al (23,25), we revisited ethanol stress by using our innovative immobilization method, which is based on trapping single yeast cells in microchambers produced by microstructured polydimethylsiloxane (PDMS) stamps (26). These microchambers can easily be loaded by convective/capillary deposition of yeast cells.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Evidence for a Role for the Plasma Membrane in the Nanomechanical Properties of the Cell Wall as Revealed by an Atomic Force Microscopy Study of the Response of Saccharomyces cerevisiae to Ethanol Stress

Schiavone

Formosa-Dague

Elsztein

et al. 2016

Appl Environ Microbiol

View full text Add to dashboard Cite

A wealth of biochemical and molecular data have been reported regarding ethanol toxicity in the yeast Saccharomyces cerevisiae. However, direct physical data on the effects of ethanol stress on yeast cells are almost nonexistent. This lack of information can now be addressed by using atomic force microscopy (AFM) technology. In this report, we show that the stiffness of glucosegrown yeast cells challenged with 9% (vol/vol) ethanol for 5 h was dramatically reduced, as shown by a 5-fold drop of Young's modulus. Quite unexpectedly, a mutant deficient in the Msn2/Msn4 transcription factor, which is known to mediate the ethanol stress response, exhibited a low level of stiffness similar to that of ethanol-treated wild-type cells. Reciprocally, the stiffness of yeast cells overexpressing MSN2 was about 35% higher than that of the wild type but was nevertheless reduced 3-to 4-fold upon exposure to ethanol. Based on these and other data presented herein, we postulated that the effect of ethanol on cell stiffness may not be mediated through Msn2/Msn4, even though this transcription factor appears to be a determinant in the nanomechanical properties of the cell wall. On the other hand, we found that as with ethanol, the treatment of yeast with the antifungal amphotericin B caused a significant reduction of cell wall stiffness. Since both this drug and ethanol are known to alter, albeit by different means, the fluidity and structure of the plasma membrane, these data led to the proposition that the cell membrane contributes to the biophysical properties of yeast cells. IMPORTANCEEthanol is the main product of yeast fermentation but is also a toxic compound for this process. Understanding the mechanism of this toxicity is of great importance for industrial applications. While most research has focused on genomic studies of ethanol tolerance, we investigated the effects of ethanol at the biophysical level and found that ethanol causes a strong reduction of the cell wall rigidity (or stiffness). We ascribed this effect to the action of ethanol perturbing the cell membrane integrity and hence proposed that the cell membrane contributes to the cell wall nanomechanical properties. T he yeast Saccharomyces cerevisiae is a remarkable ethanol producer that is also very sensitive to its main fermentative product. At low to moderate concentrations (5 to 7%), ethanol mainly affects the growth rate, and at higher concentrations (Ͼ10%), it can strongly impair cell integrity, eventually leading to cell death with features of apoptosis (1). These inhibitory and toxic effects are ascribed to the fact that ethanol alters cell membrane fluidity and dissipates the transmembrane electrochemical potential, thereby creating permeability to ionic species and causing leakage of metabolites (2). Recent works using lipidomic methodologies confirmed the relationship between the composition of lipids, notably ergosterol and unsaturated fatty acids, and ethanol tolerance (3, 4). Moreover, as it diffuses freely into cells, ethanol at high concen...

show abstract

Section: Discussioncontrasting

confidence: 57%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Evidence for a Role for the Plasma Membrane in the Nanomechanical Properties of the Cell Wall as Revealed by an Atomic Force Microscopy Study of the Response of Saccharomyces cerevisiae to Ethanol Stress

Schiavone

Formosa-Dague

Elsztein

et al. 2016

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…Saccharomyces cerevisiae, also called baker's yeast, is the best-characterized eukaryotic model for scientific and biomedical research. Although the chemical composition of the yeast cell wall is well known, its molecular ultrastructure (organization or assembly) has not been extensively studied at nanoscale (4,5), although there are a few reports on the nanomechanical and adhesive properties of the yeast cell wall under native conditions or under stress conditions (6)(7)(8). As for Candida albicans, it is by far the most common human-pathogenic fungal species.…”

mentioning

confidence: 99%

Nanoscale Effects of Caspofungin against Two Yeast Species, Saccharomyces cerevisiae and Candida albicans

Formosa

Schiavone

Martin-Yken

et al. 2013

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Saccharomyces cerevisiae and Candida albicans are model yeasts for biotechnology and human health, respectively. We used atomic force microscopy (AFM) to explore the effects of caspofungin, an antifungal drug used in hospitals, on these two species. Our nanoscale investigation revealed similar, but also different, behaviors of the two yeasts in response to treatment with the drug. While administration of caspofungin induced deep cell wall remodeling in both yeast species, as evidenced by a dramatic increase in chitin and decrease in ␤-glucan content, changes in cell wall composition were more pronounced with C. albicans cells. Notably, the increase of chitin was proportional to the increase in the caspofungin dose. In addition, the Young modulus of the cell was three times lower for C. albicans cells than for S. cerevisiae cells and increased proportionally with the increase of chitin, suggesting differences in the molecular organization of the cell wall between the two yeast species. Also, at a low dose of caspofungin (i.e., 0.5؋ MIC), the cell surface of C. albicans exhibited a morphology that was reminiscent of cells expressing adhesion proteins. Interestingly, this morphology was lost at high doses of the drug (i.e., 4؋ MIC). However, the treatment of S. cerevisiae cells with high doses of caspofungin resulted in impairment of cytokinesis. Altogether, the use of AFM for investigating the effects of antifungal drugs is relevant in nanomedicine, as it should help in understanding their mechanisms of action on fungal cells, as well as unraveling unexpected effects on cell division and fungal adhesion.T he yeast cell wall is composed of 50 to 60% ␤-glucans (glucose residues attached by 1,3-␤-and 1,6-␤-linkages), 40 to 50% mannoproteins (highly glycosylated polypeptides), and 1 to 3% chitin (1, 2). It is an essential dynamic structure playing roles in maintaining cell shape and integrity, sensing the surrounding environment, and interacting with surfaces and other cells (3). The cell wall represents 15 to 25% of the cell dry mass, the chemical composition of which is well established. Saccharomyces cerevisiae, also called baker's yeast, is the best-characterized eukaryotic model for scientific and biomedical research. Although the chemical composition of the yeast cell wall is well known, its molecular ultrastructure (organization or assembly) has not been extensively studied at nanoscale (4, 5), although there are a few reports on the nanomechanical and adhesive properties of the yeast cell wall under native conditions or under stress conditions (6-8). As for Candida albicans, it is by far the most common human-pathogenic fungal species. It can cause a range of pathogenic effects, including painful superficial infections, severe surface infections, and lifethreatening bloodstream infections (9). It is a major cause of morbidity and mortality in immunocompromised patients as a result of AIDS, cancer chemotherapy, or organ transplantation (10).Given its medical relevance, C. albicans has been the subject of extens...

show abstract

“…Ethanol concentration can reach 10% in higher gravity fermentations, and acts especially upon biological membranes [35]. Reports showed ethanol effects in growth inhibition [45], lipid modification and loss of proton motive force across the membrane and increased membrane permeability/ fluidity [46]. Yet, cells exposed to oxygen, with high levels of sterols in membranes, and adequate levels of nutrients, amino acids and trace elements in the fermentation broth are able to respond efficiently to such effects [35].…”

Section: Yeast Physiologymentioning

confidence: 99%

Yeast: World’s Finest Chef

Faria-Oliveira¹,

Puga²,

Ferreira³

2013

Food Industry

View full text Add to dashboard Cite

Atomic force microscopic study of the effects of ethanol on yeast cell surface morphology

Cited by 58 publications

References 29 publications

Evidence for a Role for the Plasma Membrane in the Nanomechanical Properties of the Cell Wall as Revealed by an Atomic Force Microscopy Study of the Response of Saccharomyces cerevisiae to Ethanol Stress

Evidence for a Role for the Plasma Membrane in the Nanomechanical Properties of the Cell Wall as Revealed by an Atomic Force Microscopy Study of the Response of Saccharomyces cerevisiae to Ethanol Stress

Nanoscale Effects of Caspofungin against Two Yeast Species, Saccharomyces cerevisiae and Candida albicans

Yeast: World’s Finest Chef

Contact Info

Product

Resources

About