The effect of oxygen concentration on the growth and metabolism of Saccharomyces cerevisiae grown with excess of potassium or in potassium-deficient media

1. Saccharomyces cerevisiae cells grown in limiting K+ concentration have their growth inhibited by O2 concentrations above 40%. With these conditions the cells grow very large and are unable to maintain ionic gradients when washed with water. 2. Cells grown in excess of K+ showed the same pattern of change in cell size with change in O2 concentration, but the magnitude of the changes was much less. Cells grown in excess of K+ were not leaky. 3. Cell death, growth and development of ‘leakiness’ were not correlated in the cells grown in limiting K+ concentration. 4. The activities of both alcohol dehydrogenase and cytochrome oxidase were higher in K+-deficient cells than in the cells grown with excess of K+. The differences were much larger when the measurements were made on a cellular basis than when made on a protein basis. 5. In 100% O2 3mm-K+ in the medium was sufficient to produce normal yeast cells.

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The Cloning and Mapping of ADR6, a Gene Required for Sporulation and for Expression of the Alcohol Dehydrogenase II Isozyme From Saccharomyces cerevisiae

Genetics ◽

10.1093/genetics/116.4.531 ◽

1987 ◽

Vol 116 (4) ◽

pp. 531-540

Author(s):

Aileen K W Taguchi ◽

Elton T Young

Keyword(s):

Saccharomyces Cerevisiae ◽

Alcohol Dehydrogenase ◽

Single Gene ◽

Carbon Sources ◽

Transcription Unit ◽

Yeast Cells ◽

Recessive Mutation ◽

Yeast Saccharomyces Cerevisiae ◽

Upstream Activating Sequence ◽

A Site

ABSTRACT The alcohol dehydrogenase II (ADH2) gene of the yeast, Saccharomyces cerevisiae, is not transcribed during growth on fermentable carbon sources such as glucose. Growth of yeast cells in a medium containing only nonfermentable carbon sources leads to a marked increase or derepression of ADH2 expression. The recessive mutation, adr6-1, leads to an inability to fully derepress ADH2 expression and to an inability to sporulate. The ADR6 gene product appears to act directly or indirectly on ADH2 sequences 3' to or including the presumptive TATAA box. The upstream activating sequence (UAS) located 5' to the TATAA box is not required for the Adr6- phenotype. Here, we describe the isolation of a recombinant plasmid containing the wild-type ADR6 gene. ADR6 codes for a 4.4-kb RNA which is present during growth both on glucose and on nonfermentable carbon sources. Disruption of the ADR6 transcription unit led to viable cells with decreased ADHII activity and an inability to sporulate. This indicates that both phenotypes result from mutations within a single gene and that the adr6-1 allele was representative of mutations at this locus. The ADR6 gene mapped to the left arm of chromosome XVI at a site 18 centimorgans from the centromere.

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Deletion of Atg22 gene contributes to reduce programmed cell death induced by acetic acid stress in Saccharomyces cerevisiae

Biotechnology for Biofuels ◽

10.1186/s13068-019-1638-x ◽

2019 ◽

Vol 12 (1) ◽

Cited By ~ 2

Author(s):

Jingjin Hu ◽

Yachen Dong ◽

Wei Wang ◽

Wei Zhang ◽

Hanghang Lou ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Amino Acids ◽

Cell Death ◽

Cell Wall ◽

Acetic Acid ◽

Programmed Cell Death ◽

Cell Wall Integrity ◽

Yeast Cells ◽

Death Process ◽

Lignocellulosic Biofuel

Abstract Background Programmed cell death (PCD) induced by acetic acid, the main by-product released during cellulosic hydrolysis, cast a cloud over lignocellulosic biofuel fermented by Saccharomyces cerevisiae and became a burning problem. Atg22p, an ignored integral membrane protein located in vacuole belongs to autophagy-related genes family; prior study recently reported that it is required for autophagic degradation and efflux of amino acids from vacuole to cytoplasm. It may alleviate the intracellular starvation of nutrition caused by Ac and increase cell tolerance. Therefore, we investigate the role of atg22 in cell death process induced by Ac in which attempt is made to discover new perspectives for better understanding of the mechanisms behind tolerance and more robust industrial strain construction. Results In this study, we compared cell growth, physiological changes in the absence and presence of Atg22p under Ac exposure conditions. It is observed that disruption and overexpression of Atg22p delays and enhances acetic acid-induced PCD, respectively. The deletion of Atg22p in S. cerevisiae maintains cell wall integrity, and protects cytomembrane integrity, fluidity and permeability upon Ac stress by changing cytomembrane phospholipids, sterols and fatty acids. More interestingly, atg22 deletion increases intracellular amino acids to aid yeast cells for tackling amino acid starvation and intracellular acidification. Further, atg22 deletion upregulates series of stress response genes expression such as heat shock protein family, cell wall integrity and autophagy. Conclusions The findings show that Atg22p possessed the new function related to cell resistance to Ac. This may help us have a deeper understanding of PCD induced by Ac and provide a new strategy to improve Ac resistance in designing industrial yeast strains for bioethanol production during lignocellulosic biofuel fermentation.

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Cell Size Influences the Reproductive Potential and Total Lifespan of theSaccharomyces cerevisiaeYeast as Revealed by the Analysis of Polyploid Strains

Oxidative Medicine and Cellular Longevity ◽

10.1155/2018/1898421 ◽

2018 ◽

Vol 2018 ◽

pp. 1-17 ◽

Cited By ~ 4

Author(s):

Renata Zadrag-Tecza ◽

Magdalena Kwolek-Mirek ◽

Małgorzata Alabrudzińska ◽

Adrianna Skoneczna

Keyword(s):

Cell Death ◽

Cell Size ◽

Reproductive Potential ◽

Threshold Value ◽

Yeast Cells ◽

Common Mechanism ◽

A Cell ◽

Genome Copy Number ◽

The Difference ◽

Two Phases

The total lifespan of the yeastSaccharomyces cerevisiaemay be divided into two phases: the reproductive phase, during which the cell undergoes mitosis cycles to produce successive buds, and the postreproductive phase, which extends from the last division to cell death. These phases may be regulated by a common mechanism or by distinct ones. In this paper, we proposed a more comprehensive approach to reveal the mechanisms that regulate both reproductive potential and total lifespan in cell size context. Our study was based on yeast cells, whose size was determined by increased genome copy number, ranging from haploid to tetraploid. Such experiments enabled us to test the hypertrophy hypothesis, which postulates that excessive size achieved by the cell—the hypertrophy state—is the reason preventing the cell from further proliferation. This hypothesis defines the reproductive potential value as the difference between the maximal size that a cell can reach and the threshold value, which allows a cell to undergo its first cell cycle and the rate of the cell size to increase per generation. Here, we showed that cell size has an important impact on not only the reproductive potential but also the total lifespan of this cell. Moreover, the maximal cell size value, which limits its reproduction capacity, can be regulated by different factors and differs depending on the strain ploidy. The achievement of excessive size by the cell (hypertrophic state) may lead to two distinct phenomena: the cessation of reproduction without “mother” cell death and the cessation of reproduction with cell death by bursting, which has not been shown before.

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Induction of apoptosis in Saccharomyces cerevisiae results in the spontaneous maturation of tetravirus procapsids in vivo

Journal of General Virology ◽

10.1099/vir.0.82250-0 ◽

2007 ◽

Vol 88 (5) ◽

pp. 1576-1582 ◽

Cited By ~ 17

Author(s):

Michele Tomasicchio ◽

Philip Arno Venter ◽

Karl H. J. Gordon ◽

Terry N. Hanzlik ◽

Rosemary Ann Dorrington

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Death ◽

Programmed Cell Death ◽

Conformational Changes ◽

Yeast Cells ◽

Genomic Rnas ◽

Virus Like Particle ◽

Induced Apoptosis

The Tetraviridae are a family of small, non-enveloped, insect RNA viruses consisting of one or two single-stranded, positive-sense genomic RNAs encapsidated in an icosahedral capsid with T=4 symmetry. Tetravirus procapsids undergo maturation when exposed to a low pH environment in vitro. While the structural biology of the conformational changes that mediate acid-dependent maturation is well understood, little is known about the significance of acid-dependent maturation in vivo. To address this question, the capsid-coding sequence of the tetravirus Helicoverpa armigera stunt virus was expressed in Saccharomyces cerevisiae cells. Virus-like particles were shown to assemble as procapsids that matured spontaneously in vivo as the cells began to age. Growth in the presence of hydrogen peroxide or acetic acid, which induced apoptosis or programmed cell death in the yeast cells, resulted in virus-like particle maturation. The results demonstrate that assembly-dependent maturation of tetravirus procapsids in vivo is linked to the onset of apoptosis in yeast cells. We propose that the reduction in pH required for tetraviral maturation may be the result of cytosolic acidification, which is associated with the early onset of programmed cell death in infected cells.

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Acrolein-Induced Oxidative Stress and Cell Death Exhibiting Features of Apoptosis in the Yeast Saccharomyces cerevisiae Deficient in SOD1

Cell Biochemistry and Biophysics ◽

10.1007/s12013-014-0376-8 ◽

2014 ◽

Vol 71 (3) ◽

pp. 1525-1536 ◽

Cited By ~ 16

Author(s):

Magdalena Kwolek-Mirek ◽

Renata Zadrąg-Tęcza ◽

Sabina Bednarska ◽

Grzegorz Bartosz

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Death ◽

Growth Arrest ◽

Yeast Cells ◽

Necrotic Cell Death ◽

Oxygen Species ◽

Necrotic Cell ◽

Yeast Saccharomyces Cerevisiae ◽

Environmental Toxin ◽

High Level

Abstract The yeast Saccharomyces cerevisiae is a useful eukaryotic model to study the toxicity of acrolein, an important environmental toxin and endogenous product of lipid peroxidation. The study was aimed at elucidation of the cytotoxic effect of acrolein on the yeast deficient in SOD1, Cu, Zn-superoxide dismutase which is hypersensitive to aldehydes. Acrolein generated within the cell from its precursor allyl alcohol caused growth arrest and cell death of the yeast cells. The growth inhibition involved an increase in production of reactive oxygen species and high level of protein carbonylation. DNA condensation and fragmentation, exposition of phosphatidylserine at the cell surface as well as decreased dynamic of actin microfilaments and mitochondria disintegration point to the induction of apoptotic-type cell death besides necrotic cell death.

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Expression of the denV gene of coliphage T4 in UV-sensitive rad mutants of Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.6.10.3559-3562.1986 ◽

1986 ◽

Vol 6 (10) ◽

pp. 3559-3562

Author(s):

K Valerie ◽

G Fronko ◽

E E Henderson ◽

J K de Riel

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Alcohol Dehydrogenase ◽

Bacteriophage T4 ◽

Yeast Cells ◽

Pyrimidine Dimer ◽

Uv Resistance ◽

Rad Mutants ◽

Cycle Dependent ◽

Denv Gene

A plasmid containing the denV gene from bacteriophage T4, under the control of the yeast alcohol dehydrogenase I (ADC1) promoter, conferred a substantial increase in UV resistance in the UV-sensitive Saccharomyces cerevisiae mutants rad1-2 and rad3-2. The UV resistance of the denV+ yeast cells was cell cycle dependent and correlated well with the level of the denV gene product as measured by immunoblotting and by a photoreversal assay for pyrimidine dimer-DNA glycosylase activity.

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Correlation of Statistical Distributions of the Dimension of Yeast Cells Attached to the Substrate and Its Surface Electrical Potential

Materials ◽

10.3390/ma15010006 ◽

2021 ◽

Vol 15 (1) ◽

pp. 6

Author(s):

Elina Bondareva ◽

Yuri Dekhtyar ◽

Vladislavs Gorosko ◽

Hermanis Sorokins ◽

Alexander Rapoport

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Adhesion ◽

Cell Size ◽

Substrate Surface ◽

Electrical Potential ◽

Statistical Distribution ◽

Yeast Cells ◽

Statistical Distributions

The ability of cells to adhere to substrates is an important factor for the effectiveness of biotechnologies and bioimplants. This research demonstrates that the statistical distribution of the sizes of the cells (Saccharomyces cerevisiae) attached to the substrate surface correlates with the statistical distribution of electrical potential on the substrate’s surface. Hypothetically, this behavior should be taken into consideration during the processing of surfaces when cell adhesion based on cell size is required.

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Characterization of the Saccharomyces cerevisiae YMR318C (ADH6) gene product as a broad specificity NADPH-dependent alcohol dehydrogenase: relevance in aldehyde reduction

Biochemical Journal ◽

10.1042/bj3610163 ◽

2001 ◽

Vol 361 (1) ◽

pp. 163-172 ◽

Cited By ~ 85

Author(s):

Carol LARROY ◽

M. Rosario FERNÁNDEZ ◽

Eva GONZÁLEZ ◽

Xavier PARÉS ◽

Josep A. BIOSCA

Keyword(s):

Saccharomyces Cerevisiae ◽

Alcohol Dehydrogenase ◽

Gene Product ◽

Yeast Cells ◽

Sequence Motif ◽

Lignin Biodegradation ◽

Primary Alcohols ◽

Medium Chain ◽

Conserved Sequence ◽

Fusel Alcohols

YMR318C represents an open reading frame from Saccharomyces cerevisiae with unknown function. It possesses a conserved sequence motif, the zinc-containing alcohol dehydrogenase (ADH) signature, specific to the medium-chain zinc-containing ADHs. In the present study, the YMR318C gene product has been purified to homogeneity from overexpressing yeast cells, and found to be a homodimeric ADH, composed of 40kDa subunits and with a pI of 5.0–5.4. The enzyme was strictly specific for NADPH and was active with a wide variety of substrates, including aliphatic (linear and branched-chain) and aromatic primary alcohols and aldehydes. Aldehydes were processed with a 50-fold higher catalytic efficiency than that for the corresponding alcohols. The highest kcat/Km values were found with pentanal>veratraldehyde > hexanal > 3-methylbutanal >cinnamaldehyde. Taking into consideration the substrate specificity and sequence characteristics of the YMR318C gene product, we have proposed this gene to be called ADH6. The disruption of ADH6 was not lethal for the yeast under laboratory conditions. Although S. cerevisiae is considered a non lignin-degrading organism, the catalytic activity of ADHVI can direct veratraldehyde and anisaldehyde, arising from the oxidation of lignocellulose by fungal lignin peroxidases, to the lignin biodegradation pathway. ADHVI is the only S. cerevisiae enzyme able to significantly reduce veratraldehyde in vivo, and its overexpression allowed yeast to grow under toxic concentrations of this aldehyde. The enzyme may also be involved in the synthesis of fusel alcohols. To our knowledge this is the first NADPH-dependent medium-chain ADH to be characterized in S. cerevisiae.

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Expression of the denV gene of coliphage T4 in UV-sensitive rad mutants of Saccharomyces cerevisiae.

Molecular and Cellular Biology ◽

10.1128/mcb.6.10.3559 ◽

1986 ◽

Vol 6 (10) ◽

pp. 3559-3562 ◽

Cited By ~ 7

Author(s):

K Valerie ◽

G Fronko ◽

E E Henderson ◽

J K de Riel

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Alcohol Dehydrogenase ◽

Bacteriophage T4 ◽

Yeast Cells ◽

Pyrimidine Dimer ◽

Uv Resistance ◽

Rad Mutants ◽

Cycle Dependent ◽

Denv Gene

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Cytochrome c Release and Mitochondria Involvement in Programmed Cell Death Induced by Acetic Acid in Saccharomyces cerevisiae

Molecular Biology of the Cell ◽

10.1091/mbc.e01-12-0161 ◽

2002 ◽

Vol 13 (8) ◽

pp. 2598-2606 ◽

Cited By ~ 261

Author(s):

Paula Ludovico ◽

Fernando Rodrigues ◽

Agostinho Almeida ◽

Manuel T. Silva ◽

Antoni Barrientos ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Death ◽

Acetic Acid ◽

Cytochrome C ◽

Programmed Cell Death ◽

Yeast Cells ◽

Cytochrome C Release ◽

Apoptotic Pathway ◽

Enzymatic Assays ◽

Species Production

Evidence is presented that mitochondria are implicated in the previously described programmed cell death (PCD) process induced by acetic acid in Saccharomyces cerevisiae. In yeast cells undergoing a PCD process induced by acetic acid, translocation of cytochrome c (CytC) to the cytosol and reactive oxygen species production, two events known to be proapoptotic in mammals, were observed. Associated with these events, reduction in oxygen consumption and in mitochondrial membrane potential was found. Enzymatic assays showed that the activity of complexbc 1 was normal, whereas that of cytochrome c oxidase (COX) was strongly decreased. This decrease is in accordance with the observed reduction in the amounts of COX II subunit and of cytochromesa+a 3 . The acetic acid-induced PCD process was found to be independent of oxidative phosphorylation because it was not inhibited by oligomycin treatment. The inability ofS. cerevisiae mutant strains (lacking mitochondrial DNA, heme lyase, or ATPase) to undergo acetic acid-induced PCD and in the ATPase mutant (knockout in ATP10) the absence of CytC release provides further evidence that the process is mediated by a mitochondria-dependent apoptotic pathway. The understanding of the involvement of a mitochondria-dependent apoptotic pathway inS. cerevisiae PCD process will be most useful in the further elucidation of an ancestral pathway common to PCD in metazoans.

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