scholarly journals Saccharomyces cerevisiae Concentrates Subtoxic Copper onto Cell Wall from Solid Media Containing Reducing Sugars as Carbon Source

2021 ◽  
Vol 8 (3) ◽  
pp. 36
Author(s):  
Lavinia L. Ruta ◽  
Ileana C. Farcasanu
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Wall ◽  
Carbon Source ◽  
Copper Concentration ◽  
Reducing Sugars ◽  
Carbon Sources ◽  
Wine Production ◽  
Physiological Limits ◽  
High Copper ◽  
Cell Wall Disruption

Copper is essential for life, but it can be deleterious in concentrations that surpass the physiological limits. Copper pollution is related to widespread human activities, such as viticulture and wine production. To unravel aspects of how organisms cope with copper insults, we used Saccharomyces cerevisiae as a model for adaptation to high but subtoxic concentrations of copper. We found that S. cerevisiae cells could tolerate high copper concentration by forming deposits on the cell wall and that the copper-containing deposits accumulated predominantly when cells were grown statically on media prepared with reducing sugars (glucose, galactose) as sole carbon source, but not on media containing nonreducing carbon sources, such as glycerol or lactate. Exposing cells to copper in liquid media under strong agitation prevented the formation of copper-containing deposits at the cell wall. Disruption of low-affinity copper intake through the plasma membrane increased the potential of the cell to form copper deposits on the cell surface. These results imply that biotechnology problems caused by high copper concentration can be tackled by selecting yeast strains and conditions to allow the removal of excess copper from various contaminated sites in the forms of solid deposits which do not penetrate the cell.

scholarly journals CAT8, a new zinc cluster-encoding gene necessary for derepression of gluconeogenic enzymes in the yeast Saccharomyces cerevisiae.

1995 ◽  
Vol 15 (4) ◽  
pp. 1915-1922 ◽  
Author(s):  
D Hedges ◽  
M Proft ◽  
K D Entian
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Kinase ◽  
Carbon Source ◽  
Gene Activation ◽  
Carbon Sources ◽  
Source Analysis ◽  
Yeast Saccharomyces Cerevisiae ◽  
Gluconeogenic Enzymes ◽  
Zinc Cluster ◽  
Promoter Fusion

The expression of gluconeogenic fructose-1,6-bisphosphatase (encoded by the FBP1 gene) depends on the carbon source. Analysis of the FBP1 promoter revealed two upstream activating elements, UAS1FBP1 and UAS2FBP1, which confer carbon source-dependent regulation on a heterologous reporter gene. On glucose media neither element was activated, whereas after transfer to ethanol a 100-fold derepression was observed. This gene activation depended on the previously identified derepression genes CAT1 (SNF1) (encoding a protein kinase) and CAT3 (SNF4) (probably encoding a subunit of Cat1p [Snf1p]). Screening for mutations specifically involved in UAS1FBP1 derepression revealed the new recessive derepression mutation cat8. The cat8 mutants also failed to derepress UAS2FBP1, and these mutants were unable to grow on nonfermentable carbon sources. The CAT8 gene encodes a zinc cluster protein related to Saccharomyces cerevisiae Gal4p. Deletion of CAT8 caused a defect in glucose derepression which affected all key gluconeogenic enzymes. Derepression of glucose-repressible invertase and maltase was still normally regulated. A CAT8-lacZ promoter fusion revealed that the CAT8 gene itself is repressed by Cat4p (Mig1p). These results suggest that gluconeogenic genes are derepressed upon binding of Cat8p, whose synthesis depends on the release of Cat4p (Mig1p) from the CAT8 promoter. However, gluconeogenic promoters are still glucose repressed in cat4 mutants, which indicates that in addition to its transcription, the Cat8p protein needs further activation. The observation that multicopy expression of CAT8 reverses the inability of cat1 and cat3 mutants to grow on ethanol indicates that Cat8p might be the substrate of the Cat1p/Cat3p protein kinase.

scholarly journals Growth optimization of Saccharomyces cerevisiae and Rhizopus oligosporus during fermentation to produce tempeh with high β-glucan content

2020 ◽  
Vol 21 (6) ◽  
Author(s):  
SAMSUL RIZAL ◽  
Murhadi Murhadi ◽  
Maria Erna Kustyawati ◽  
Udin Hasanudin
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Wheat Flour ◽  
Ph Value ◽  
Block Design ◽  
Carbon Sources ◽  
Rhizopus Oligosporus ◽  
Growth Optimization ◽  
Yeast Fungi ◽  
Two Factors

Abstract. Rizal S, Murhadi, Kustyawati ME, Hasanudin U. 2020. Growth optimization of Saccharomyces cerevisiae and Rhizopus oligosporus during fermentation to produce tempeh with high β-glucan content. Biodiversitas 21: 2667-2673. Saccharomyces cerevisiae grows and produces β-glucan during fermentation in tempeh production. The content of β-glucan in tempeh is influenced by the growth of S. cerevisiae throughout fermentation. The purpose of this study was to determine the effects of different types and concentrations of carbon sources on yeast growth, fungi growth, and β-glucan content in tempeh inoculated using Rhizopus oligosporus and S. cerevisiae. This study used a Factorial Randomized Complete Block Design (RCBD) with two factors and three replications. The first factor was the types of carbon sources, tapioca and wheat flour; the second factor was the concentrations of carbon source, 0.0%, 2.5%, 5.0%, 7.5% and 10.0% (w/w). Tempeh produced was investigated for yeast number, fungi number, β-glucan content, and pH value. The obtained data were tested using Tukey's Honestly Significance Difference (HSD) test. The results showed that the addition of various types and concentrations of carbon source significantly influenced the increase in yeast number, fungi number, β-glucan content, and pH in tempeh. The growth of yeast, fungi, and β-glucan content increased along with the increment of carbon source concentration. The amounts of yeast, fungi, and β-glucans in tempeh added with tapioca were higher compared to tempeh with wheat flour. The addition of 10% tapioca produced the highest amount of yeast with 9.505 Log CFU/g and the highest β-glucan content with 0.707% (w/w).

scholarly journals A common bacterial metabolite elicits prion-based bypass of glucose repression

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
David M Garcia ◽  
David Dietrich ◽  
Jon Clardy ◽  
Daniel F Jarosz
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Lactic Acid ◽  
Glucose Metabolism ◽  
Carbon Source ◽  
Glucose Repression ◽  
Carbon Sources ◽  
Yeast Cells ◽  
Genetic Element ◽  
Yeast Saccharomyces Cerevisiae ◽  
The Common

Robust preference for fermentative glucose metabolism has motivated domestication of the budding yeast Saccharomyces cerevisiae. This program can be circumvented by a protein-based genetic element, the [GAR+] prion, permitting simultaneous metabolism of glucose and other carbon sources. Diverse bacteria can elicit yeast cells to acquire [GAR+], although the molecular details of this interaction remain unknown. Here we identify the common bacterial metabolite lactic acid as a strong [GAR+] inducer. Transient exposure to lactic acid caused yeast cells to heritably circumvent glucose repression. This trait had the defining genetic properties of [GAR+], and did not require utilization of lactic acid as a carbon source. Lactic acid also induced [GAR+]-like epigenetic states in fungi that diverged from S. cerevisiae ~200 million years ago, and in which glucose repression evolved independently. To our knowledge, this is the first study to uncover a bacterial metabolite with the capacity to potently induce a prion.

ABF1 is a phosphoprotein and plays a role in carbon source control of COX6 transcription in Saccharomyces cerevisiae

1992 ◽  
Vol 12 (9) ◽  
pp. 4197-4208
Author(s):  
S Silve ◽  
P R Rhode ◽  
B Coll ◽  
J Campbell ◽  
R O Poyton
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Glucose Repression ◽  
Carbon Sources ◽  
Growth Conditions ◽  
Source Control ◽  
Specific Target Gene ◽  
Phosphorylation Pattern ◽  
Nonfermentable Carbon Source ◽  
In Cells

Previously, we have shown that the Saccharomyces cerevisiae DNA-binding protein ABF1 exists in at least two different electrophoretic forms (K. S. Sweder, P. R. Rhode, and J. L. Campbell, J. Biol. Chem. 263: 17270-17277, 1988). In this report, we show that these forms represent different states of phosphorylation of ABF1 and that at least four different phosphorylation states can be resolved electrophoretically. The ratios of these states to one another differ according to growth conditions and carbon source. Phosphorylation of ABF1 is therefore a regulated process. In nitrogen-starved cells or in cells grown on nonfermentable carbon sources (e.g., lactate), phosphorylated forms predominate, while in cells grown on fermentable carbon sources (e.g., glucose), dephosphorylated forms are enriched. The phosphorylation pattern is affected by mutations in the SNF1-SSN6 pathway, which is involved in glucose repression-depression. Whereas a functional SNF1 gene, which encodes a protein kinase, is not required for the phosphorylation of ABF1, a functional SSN6 gene is required for itsd ephosphorylation. The phosphorylation patterns that we have observed correlate with the regulation of a specific target gene, COX6, which encodes subunit VI of cytochrome c oxidase. Transcription of COX6 is repressed by growth in medium containing a fermentable carbon source and is derepressed by growth in medium containing a nonfermentable carbon source. COX6 repression-derepression is under the control of the SNF1-SSN6 pathway. This carbon source regulation is exerted through domain 1, a region of the upstream activation sequence UAS6 that binds ABF1 (J. D. Trawick, N. Kraut, F. Simon, and R. O. Poyton, Mol. Cell Biol. 12:2302-2314, 1992). We show that the greater the phosphorylation of ABF1, the greater the transcription of COX6. Furthermore, the ABF1-containing protein-DNA complexes formed at domain 1 differ according to the phosphorylation state of ABF1 and the carbon source on which the cells were grown. From these findings, we propose that the phosphorylation of ABF1 is involved in glucose repression-derepression of COX6 transcription.

Saccharomyces cerevisiae glucose signalling regulator Mth1p regulates the organellar Na+/H+ exchanger Nhx1p

2010 ◽  
Vol 432 (2) ◽  
pp. 343-352 ◽  
Author(s):  
Keiji Mitsui ◽  
Masafumi Matsushita ◽  
Hiroshi Kanazawa
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Sole Carbon Source ◽  
Glucose Sensor ◽  
Ph Regulation ◽  
Gene Knockout ◽  
Carbon Sources ◽  
Acidic Ph ◽  
In Cells

Organelle-localized NHEs (Na+/H+ exchangers) are found in cells from yeast to humans and contribute to organellar pH regulation by exporting H+ from the lumen to the cytosol coupled to an H+ gradient established by vacuolar H+-ATPase. The mechanisms underlying the regulation of organellar NHEs are largely unknown. In the present study, a yeast two-hybrid assay identified Mth1p as a new binding protein for Nhx1p, an organellar NHE in Saccharomyces cerevisiae. It was shown by an in vitro pull-down assay that Mth1p bound to the hydrophilic C-terminal half of Nhx1p, especially to the central portion of this region. Mth1p is known to bind to the cytoplasmic domain of the glucose sensor Snf3p/Rgt2p and also functions as a negative transcriptional regulator. Mth1p was expressed in cells grown in a medium containing galactose, but was lost (possibly degraded) when cells were grown in medium containing glucose as the sole carbon source. Deletion of the MTH1 gene increased cell growth compared with the wild-type when cells were grown in a medium containing galactose and with hygromycin or at an acidic pH. This resistance to hygromycin or acidic conditions was not observed for cells grown with glucose as the sole carbon source. Gene knockout of NHX1 increased the sensitivity to hygromycin and acidic pH. The increased resistance to hygromycin was reproduced by truncation of the Mth1p-binding region in Nhx1p. These results implicate Mth1p as a novel regulator of Nhx1p that responds to specific extracellular carbon sources.

scholarly journals SDC25, a dispensable Ras guanine nucleotide exchange factor of Saccharomyces cerevisiae differs from CDC25 by its regulation.

1996 ◽  
Vol 7 (4) ◽  
pp. 529-539 ◽  
Author(s):  
E Boy-Marcotte ◽  
P Ikonomi ◽  
M Jacquet
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Transcriptional Control ◽  
Carbon Sources ◽  
Guanine Nucleotide Exchange Factor ◽  
Guanine Nucleotide ◽  
Nucleotide Exchange ◽  
Exchange Factor ◽  
Guanine Nucleotide Exchange ◽  
Nucleotide Exchange Factor

The SDC25 gene of Saccharomyces cerevisiae is homologous to CDC25. Its 3' domain encodes a guanine nucleotide exchange factor (GEF) for Ras. Nevertheless, the GEF encoded by CDC24 is determinant for the Ras/cAMP pathway activation in growth. We demonstrate that the SDC25 gene product is a functional GEF for Ras: the complete SDC25 gene functionally replaces CDC25 when overexpressed or when transcribed under CDC25 transcriptional control at the CDC25 locus. Chimeric proteins between Sdc25p and Cdc25p are also functional GEFs for Ras. We also show that the two genes are differentially regulated: SDC25 is not transcribed at a detectable level in growth conditions when glucose is the carbon source. It is transcribed at the end of growth when nutrients are depleted and in cells grown on nonfermentable carbon sources. In contrast, CDC25 accumulation is slightly reduced when glucose is replaced by a nonfermentable carbon source.

scholarly journals The Function and Properties of the Azf1 Transcriptional Regulator Change with Growth Conditions in Saccharomyces cerevisiae

2006 ◽  
Vol 5 (2) ◽  
pp. 313-320 ◽  
Author(s):  
Matthew G. Slattery ◽  
Dritan Liko ◽  
Warren Heideman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Wall ◽  
Quaternary Structure ◽  
Protein A ◽  
Sodium Dodecyl ◽  
Carbon Sources ◽  
Extraction Procedure ◽  
Growth Conditions ◽  
Growth Defect ◽  
Calcofluor White

ABSTRACT Azf1 activates CLN3 transcription in Saccharomyces cerevisiae cells growing in glucose. Paradoxically, other studies have shown Azf1 to be almost undetectable in glucose-grown cells. Microarray experiments showed that Azf1 activates nonoverlapping gene sets in different carbon sources: in glucose, Azf1 activates carbon and energy metabolism genes, and in glycerol-lactate, Azf1 activates genes needed for cell wall maintenance. Consistent with the decreased expression of cell wall maintenance genes observed with azf1Δ mutants, we observed a marked growth defect in the azf1Δ cells at 37°C in nonfermentable medium. Cell wall integrity assays, such as sensitivity to calcofluor white, sodium dodecyl sulfate, or caffeine, confirmed cell wall defects in azf1Δ mutants in nonfermentable medium. Gel shift experiments show that Azf1 binds to DNA elements with the sequence AAAAGAAA (A4GA3), a motif enriched in the promoters of Azf1-sensitive genes and predicted by whole-genome studies. This suggests that many of the Azf1-dependent transcripts may be regulated directly by Azf1 binding. We found that the levels of Azf1 protein in glucose-grown cells were comparable to Azf1 levels in cells grown in glycerol-lactate; however, this could only be demonstrated with a cell extraction procedure that minimizes proteolysis. Glucose produces conditions that destabilize the Azf1 protein, a finding that may reflect a glucose-induced change in Azf1 tertiary or quaternary structure.

scholarly journals Aca1 and Aca2, ATF/CREB Activators in Saccharomyces cerevisiae, Are Important for Carbon Source Utilization but Not the Response to Stress

2000 ◽  
Vol 20 (12) ◽  
pp. 4340-4349 ◽  
Author(s):  
M. Adelaida Garcia-Gimeno ◽  
Kevin Struhl
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Target Genes ◽  
Carbon Sources ◽  
Biological Functions ◽  
Carbon Source Utilization ◽  
The Family ◽  
Response To Stress

ABSTRACT In Saccharomyces cerevisiae, the family of ATF/CREB transcriptional regulators consists of a repressor, Acr1 (Sko1), and two activators, Aca1 and Aca2. The AP-1 factor Gen4 does not activate transcription through ATF/CREB sites in vivo even though it binds these sites in vitro. Unlike ATF/CREB activators in other species, Aca1- and Aca2-dependent transcription is not affected by protein kinase A or by stress, and Aca1 and Aca2 are not required for Hog1-dependent salt induction of transcription through an optimal ATF/CREB site. Aca2 is important for a variety of biological functions including growth on nonoptimal carbon sources, and Aca2-dependent activation is modestly regulated by carbon source. Strains lacking Aca1 are phenotypically normal, but overexpression of Aca1 suppresses some defects associated with the loss of Aca2, indicating a functional overlap between Aca1 and Aca2. Acr1 represses transcription both by recruiting the Cyc8-Tup1 corepressor and by directly competing with Aca1 and Aca2 for target sites. Acr1 does not fully account for osmotic regulation through ATF/CREB sites, and a novel Hog1-dependent activator(s) that is not a bZIP protein is required for ATF/CREB site activation in response to high salt. In addition, Acr1 does not affect a number of phenotypes that arise from loss of Aca2. Thus, members of the S. cerevisiae ATF/CREB family have overlapping, but distinct, biological functions and target genes.

scholarly journals The Defect in Transcription-Coupled Repair Displayed by a Saccharomyces cerevisiae rad26 Mutant Is Dependent on Carbon Source and Is Not Associated With a Lack of Transcription

Genetics ◽  
2001 ◽  
Vol 158 (3) ◽  
pp. 989-997
Author(s):  
Miriam Bucheli ◽  
Lori Lommel ◽  
Kevin Sweder
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Damage ◽  
Carbon Source ◽  
Excision Repair ◽  
Carbon Sources ◽  
Repair Process ◽  
Growth Conditions ◽  
Nucleotide Excision ◽  
Evolutionarily Conserved ◽  
Transcription Activators

Abstract Nucleotide excision repair (NER) is an evolutionarily conserved pathway that removes DNA damage induced by ultraviolet irradiation and various chemical agents that cause bulky adducts. Two subpathways within NER remove damage from the genome overall or the transcribed strands of transcribing genes (TCR). TCR is a faster repair process than overall genomic repair and has been thought to require the RAD26 gene in Saccharomyces cerevisiae. Rad26 is a member of the SWI/SNF family of proteins that either disrupt chromatin or facilitate interactions between the RNA Pol II and transcription activators. SWI/SNF proteins are required for the expression or repression of a diverse set of genes, many of which are differentially transcribed in response to particular carbon sources. The remodeling of chromatin by Rad26 could affect transcription and/or TCR following formation of DNA damage and other stress-inducing conditions. We speculate that another factor(s) can substitute for Rad26 under particular growth conditions. We therefore measured the level of repair and transcription in two different carbon sources and found that the defect in the rad26 mutant for TCR was dependent on the type of carbon source. Furthermore, TCR did not correlate with transcription rate, suggesting that disruption of RAD26 leads to a specific defect in DNA repair and not transcription.

scholarly journals Accelerated Alcoholic Fermentation of Intact Grapes by Saccharomyces Cerevisiae in Symbiosis with Microbial Community Inhabiting Grape-skin

2021 ◽  
Author(s):  
Daisuke Watanabe ◽  
Wataru Hashimoto
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Sole Carbon Source ◽  
Alcoholic Fermentation ◽  
Carbon Sources ◽  
Grape Skin ◽  
Carbon Availability ◽  
Grape Berries ◽  
Symbiotic Interactions ◽  
Fungal Microbiota

Abstract Saccharomyces cerevisiae, an essential player in alcoholic fermentation during winemaking, is rarely found in intact grapes. Here, we addressed symbiotic interactions between S. cerevisiae and grape-skin residents upon spontaneous wine fermentation. When glucose was used as a carbon source, the yeast-like fungus Aureobasidium pullulans, a major grape-skin resident, had no effect on alcoholic fermentation by S. cerevisiae. In contrast, when intact grape berries as a sole carbon source, coculture of S. cerevisiae and A. pullulans accelerated alcoholic fermentation. Thus, grape-inhabiting microorganisms may increase carbon availability by degrading and/or incorporating grape-skin materials, such as cell wall and cuticles. A. pullulans exhibited broad spectrum assimilation of plant-derived carbon sources, including ω-hydroxy fatty acids, arising from degradation of cutin. In fact, yeast-type cutinase was produced from A. pullulans EXF-150 strain. The degradation and utilization of grape-skin materials by fungal microbiota may account for their colonization on grape-skin and symbiotic interactions with S. cerevisiae.

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