Kcs1 and Vip1: The Key Enzymes behind Inositol Pyrophosphate Signaling in Saccharomyces cerevisiae.

The inositol pyrophosphate pathway, a complex cell signaling network, plays a pivotal role in orchestrating vital cellular processes in the budding yeast, where it regulates cell cycle progression, growth, endocytosis, exocytosis, apoptosis, telomere elongation, ribosome biogenesis, and stress responses. This pathway has gained significant attention in pharmacology and medicine due to its role in generating inositol pyrophosphates, which serve as crucial signaling molecules not only in yeast, but also in higher eukaryotes. As targets for therapeutic development, genetic modifications within this pathway hold promise for disease treatment strategies, offering practical applications in biotechnology. The model organism Saccharomyces cerevisiae, renowned for its genetic tractability, has been instrumental in various studies related to the inositol pyrophosphate pathway. This review is focused on the Kcs1 and Vip1, the two enzymes involved in the biosynthesis of inositol pyrophosphate in S. cerevisiae, highlighting their roles in various cell processes, and providing an up-to-date overview of their relationship with phosphate homeostasis. Moreover, the review underscores the potential applications of these findings in the realms of medicine and biotechnology, highlighting the profound implications of comprehending this intricate signaling network.

New Heterocyclic Compounds from Oxazol-5(4H)-one and 1,2,4-Triazin-6(5H)-one Classes: Synthesis, Characterization and Toxicity Evaluation.

This paper describes the synthesis of new heterocycles from oxazol-5(4H)-one and 1,2,4-triazin-6(5H)-one classes containing a phenyl-/4-bromophenylsulfonylphenyl moiety. The oxazol-5(4H)-ones were obtained via condensation of 2-(4-(4-X-phenylsulfonyl)benzamido)acetic acids with benzaldehyde/4-fluorobenzaldehyde in acetic anhydride and in the presence of sodium acetate. The reaction of oxazolones with phenylhydrazine, in acetic acid and sodium acetate, yielded the corresponding 1,2,4-triazin-6(5H)-ones. The structures of the compounds were confirmed using spectral (FT-IR, 1H-NMR, 13C-NMR, MS) and elemental analysis. The toxicity of the compounds was evaluated on Daphnia magna Straus crustaceans and on the budding yeast Saccharomyces cerevisiae. The results indicate that both the heterocyclic nucleus and halogen atoms significantly influenced the toxicity against D. magna, with the oxazolones being less toxic than triazinones. The halogen-free oxazolone had the lowest toxicity, and the fluorine-containing triazinone exhibited the highest toxicity. The compounds showed low toxicity against yeast cells, apparently due to the activity of plasma membrane multidrug transporters Pdr5 and Snq2. The predictive analyses indicated an antiproliferative effect as the most probable biological action. The PASS prediction and CHEMBL similarity studies show evidence that the compounds could inhibit certain relevant oncological protein kinases. These results correlated with toxicity assays suggest that halogen-free oxazolone could be a good candidate for future anticancer investigations.

Interaction between Polyphenolic Antioxidants and Saccharomyces cerevisiae Cells Defective in Heavy Metal Transport across the Plasma Membrane.

Natural polyphenols are compounds with important biological implications which include antioxidant and metal-chelating characteristics relevant for their antimicrobial, antitumor, or antiaging potential. The mechanisms linking polyphenols and heavy metals in their concerted actions on cells are not completely elucidated. In this study, we used the model eukaryotic microorganism Saccharomyces cerevisiae to detect the action of widely prevalent natural polyphenols on yeast cells defective in the main components involved in essential heavy metal transport across the plasma membrane. We found that caffeic and gallic acids interfered with Zn accumulation, causing delays in cell growth that were alleviated by Zn supplementation. The flavones morin and quercetin interfered with both Mn and Zn accumulation, which resulted in growth improvement, but supplemental Mn and especially Zn turned the initially benefic action of morin and quercetin into potential toxicity. Our results imply that caution is needed when administering food supplements or nutraceuticals which contain both natural polyphenols and essential elements, especially zinc.

Saccharomyces cerevisiae cells lacking transcription factors Skn7 or Yap1 exhibit different susceptibility to cyanidin.

Anthocyanidins - the aglycone moiety of anthocyanins - are responsible for the antioxidant traits and for many of the health benefits brought by the consumption of anthocyanin-rich foods, but whether excessive anthocyanidins are deleterious to living organisms is still a matter of debate. In the present study we used the model eukaryotic microorganism Saccharomyces cerevisiae to evaluate the potential toxicity of cyanidin, one of the most prevalent anthocyanidins found in berries, grapes, purple vegetables, and red wine. We found that yeast cells lacking the transcription factors responsible for regulating the response to oxidative stress - Skn7 and Yap1 - exhibited different sensitivities to cyanidin. Cells lacking the transcription factor Skn7 were sensitive to low concentrations of cyanidin, a trait that was augmented by exposure to visible light, notably blue or green light. In contrast, the growth of yeast cells devoid of Yap1 was stimulated by low concentrations, but it was impaired by high cyanidin exposure. High, but not low cyanidin was shown to induce Yap1 translocation from cytosol to nucleus, probably by generating reactive oxygen species such as H2O2. Taken together, these observation suggested that Skn7 and Yap1 have complementary roles in adaptation to cyanidin stress, with Skn7 involved in adaptation to low concentrations and with Yap1 responsible for adaptation to high concentrations of cyanidin. The results imply that caution is needed when utilizing cyanidin-enriched supplements, especially when in combination with prolonged exposure to visible light.

Saccharomyces cerevisiae and Caffeine Implications on the Eukaryotic Cell.

Caffeine-a methylxanthine analogue of the purine bases adenine and guanine-is by far the most consumed neuro-stimulant, being the active principle of widely consumed beverages such as coffee, tea, hot chocolate, and cola. While the best-known action of caffeine is to prevent sleepiness by blocking the adenosine receptors, caffeine exerts a pleiotropic effect on cells, which lead to the activation or inhibition of various cell integrity pathways. The aim of this review is to present the main studies set to investigate the effects of caffeine on cells using the model eukaryotic microorganism Saccharomyces cerevisiae, highlighting the caffeine synergy with external cell stressors, such as irradiation or exposure to various chemical hazards, including cigarette smoke or chemical carcinogens. The review also focuses on the importance of caffeine-related yeast phenotypes used to resolve molecular mechanisms involved in cell signaling through conserved pathways, such as target of rapamycin (TOR) signaling, Pkc1-Mpk1 mitogen activated protein kinase (MAPK) cascade, or Ras/cAMP protein kinase A (PKA) pathway.

Anthocyanins and Anthocyanin-Derived Products in Yeast-Fermented Beverages.

The beverages obtained by yeast fermentation from anthocyanin-rich natural sources (grapes, berries, brown rice, etc.) retain part of the initial pigments in the maturated drink. During the fermentation and aging processes anthocyanins undergo various chemical transformations, which include reactions with glycolytic products (especially pyruvate and acetaldehyde) or with other compounds present in the complex fermentation milieu (such as vinylphenols obtained from cinnamic acids by means of a yeast decarboxylase) yielding pigments which can be more stable than the initial anthocyanins. Overall, these compounds contribute to the organoleptic traits of the mature product, but also to the overall chemical composition which make the yeast fermented beverages important sources of dietary antioxidants. In this review, we focused on the studies regarding the changes underwent by anthocyanins during yeast-mediated fermentation, on the approaches taken to enrich the fermented beverages in anthocyanins and their derived products, and on the interrelations between yeast and anthocyanin which were of relevance for obtaining a high-quality product containing optimum amounts of anthocyanin and anthocyanin-derived products.

Anchoring plant metallothioneins to the inner face of the plasma membrane of Saccharomyces cerevisiae cells leads to heavy metal accumulation.

In this study we engineered yeast cells armed for heavy metal accumulation by targeting plant metallothioneins to the inner face of the yeast plasma membrane. Metallothioneins (MTs) are cysteine-rich proteins involved in the buffering of excess metal ions, especially Cu(I), Zn(II) or Cd(II). The cDNAs of seven Arabidopsis thaliana MTs (AtMT1a, AtMT1c, AtMT2a, AtMT2b, AtMT3, AtMT4a and AtMT4b) and four Noccaea caerulescens MTs (NcMT1, NcMT2a, NcMT2b and NcMT3) were each translationally fused to the C-terminus of a myristoylation green fluorescent protein variant (myrGFP) and expressed in Saccharomyces cerevisiae cells. The myrGFP cassette introduced a yeast myristoylation sequence which allowed directional targeting to the cytosolic face of the plasma membrane along with direct monitoring of the intracellular localization of the recombinant protein by fluorescence microscopy. The yeast strains expressing plant MTs were investigated against an array of heavy metals in order to identify strains which exhibit the (hyper)accumulation phenotype without developing toxicity symptoms. Among the transgenic strains which could accumulate Cu(II), Zn(II) or Cd(II), but also non-canonical metal ions, such as Co(II), Mn(II) or Ni(II), myrGFP-NcMT3 qualified as the best candidate for bioremediation applications, thanks to the robust growth accompanied by significant accumulative capacity.

Heavy metal accumulation by Saccharomyces cerevisiae cells armed with metal binding hexapeptides targeted to the inner face of the plasma membrane.

Accumulation of heavy metals without developing toxicity symptoms is a phenotype restricted to a small group of plants called hyperaccumulators, whose metal-related characteristics suggested the high potential in biotechnologies such as bioremediation and bioextraction. In an attempt to extrapolate the heavy metal hyperaccumulating phenotype to yeast, we obtained Saccharomyces cerevisiae cells armed with non-natural metal-binding hexapeptides targeted to the inner face of the plasma membrane, expected to sequester the metal ions once they penetrated the cell. We describe the construction of S. cerevisiae strains overexpressing metal-binding hexapeptides (MeBHxP) fused to the carboxy-terminus of a myristoylated green fluorescent protein (myrGFP). Three non-toxic myrGFP-MeBHxP (myrGFP-H6, myrGFP-C6, and myrGFP-(DE)3) were investigated against an array of heavy metals in terms of their effect on S. cerevisiae growth, heavy metal (hyper) accumulation, and capacity to remove heavy metal from contaminated environments.

Heat shock, visible light or high calcium augment the cytotoxic effects of Ailanthus altissima (Swingle) leaf extracts against Saccharomyces cerevisiae cells.

To gain new insight into the antimicrobial potential of Ailanthus altissima Swingle, ethanol leaf extracts were evaluated for the antifungal effects against the model yeast Saccharomyces cerevisae. The extracts inhibited the yeast growth in a dose-dependent manner, and this effect could be augmented by heat shock, exposure to visible light or exposure to high concentrations of Ca(2+). Using transgenic yeast cells expressing the Ca(2+)-dependent photoprotein, aequorin, it was found that the leaf extracts induced cytosolic Ca(2+) elevation. Experiments on yeast mutants with defects in Ca(2+) transport demonstrated that the cytotoxicity of the A. altissima leaf extracts (AaLEs) was mediated by transient pulses of Ca(2+) ions which were released into the cytosol predominantly from the vacuole. The investigation of the antifungal synergies involving AaLEs may contribute to the development of optimal and safe combination therapies for the treatment of drug-resistant fungal infections.

Overexpression of the PHO84 gene causes heavy metal accumulation and induces Ire1p-dependent unfolded protein response in Saccharomyces cerevisiae cells.

Pho84p, the protein responsible for the high-affinity uptake and transport of inorganic phosphate across the plasma membrane, is also involved in the low-affinity uptake of heavy metals in the Saccharomyces cerevisiae cells. In the present study, the effect of PHO84 overexpression upon the heavy metal accumulation by yeast cells was investigated. As PHO84 overexpression triggered the Ire1p-dependent unfolded protein response, abundant plasma membrane Pho84p could be achieved only in ire1Δ cells. Under environmental surplus, PHO84 overexpression augmented the metal accumulation by the wild type, accumulation that was exacerbated by the IRE1 deletion. The pmr1Δ cells, lacking the gene that encodes the P-type ATPase ion pump that transports Ca(2+) and Mn(2+) into the Golgi, hyperaccumulated Mn(2+) even from normal medium when overexpressing PHO84, a phenotype which is rather restricted to metal-hyperaccumulating plants.

Removing heavy metals from synthetic effluents using "kamikaze" Saccharomyces cerevisiae cells.

One key step of the bioremediation processes designed to clean up heavy metal contaminated environments is growing resistant cells that accumulate the heavy metals to ensure better removal through a combination of biosorption and continuous metabolic uptake after physical adsorption. Saccharomyces cerevisiae cells can easily act as cation biosorbents, but isolation of mutants that are both hyperaccumulating and tolerant to heavy metals proved extremely difficult. Instead, mutants that are hypersensitive to heavy metals due to increased and continuous uptake from the environment were considered, aiming to use such mutants to reduce the heavy metal content of contaminated waters. In this study, the heavy metal hypersensitive yeast strain pmr1Delta was investigated for the ability to remove Mn2+, Cu2+, Co2+, or Cd2+ from synthetic effluents. Due to increased metal accumulation, the mutant strain was more efficient than the wild-type in removing Mn2+, Cu2+, or Co2+ from synthetic effluents containing 1-2 mM cations, with a selectivity and also in removing Mn2+ and Cd2+ from synthetic effluents containing 20-50 microM cations, with a selectivity Mn2+ > Cd2+.

The antioxidant response induced by Lonicera caerulaea berry extracts in animals bearing experimental solid tumors.

Lonicera caerulea is a species of bush native to the Kamchatka Peninsula (Russian Far East) whose berries have been extensively studied due to their potential high antioxidant activity. The aim of our work was to investigate the in vivo effects of the antioxidant action of Lonicera caerulea berry extracts on the dynamics of experimentally-induced tumors. Our data showed that aqueous Lonicera caerulaea extracts reduced the tumor volume when administered continuously during the tumor growth and development stages, but augmented the tumor growth when the administration of extracts started three weeks before tumor grafting. Prolonged administration of Lonicera caerulaea berry extracts induced the antioxidant defense mechanism in the tumor tissues, while surprisingly amplifying the peripheral oxidative stress.

Chemical and biological studies of Ribes nigrum L. buds essential oil.

Ribes nigrum buds are used in medicine for the diuretic and antiseptic properties of their volatile compounds. We present in this paper comparative data concerning the chemical composition of Ribes nigrum buds essential oils obtained from three blackcurrant varieties. Essential oils were isolated by steam distillation and were analyzed by gas chromatography coupled with mass spectrometry. The Ribes nigrum essential oils (extracted from all three varieties) exhibited similar and large antibacterial spectrum, acting against Acinetobacter (A.) baumanii, Escherichia (E.) coli, Pseudomonas (P.) aeruginoasa and Staphylococcus (S.) aureus, as proved by the very low MIC values observed for the respective strains. The subinhibitory concentrations of the essential oils induced a decrease in the bacterial ability to colonize the inert substratum for A. baumanii, E. coli and S. aureus, demonstrating that besides the bactericidal activity, the Ribes nigrum essential oils also exhibit anti-pathogenic potential.

Role of L-histidine in conferring tolerance to Ni2+ in Sacchromyces cerevisiae cells.

Ni(2+) toxicity can be alleviated in yeast cells by exogenous L-histidine, but not by its enantiomer, D-histidine, nor by other natural L-amino acids tested. We studied the effect of L-histidine upon the accumulation and intracellular distribution of Ni(2+) and found that moderate L-histidine concentrations (less than or equal to those of Ni(2+)) increased cell tolerance without decreasing Ni(2+) accumulation. Although excess L-histidine appeared to lower Ni(2+) accumulation, the concomitant presence of Ni(2+) and L-histidine in the growth medium stimulated each other's uptake.