Effects of a Heterogenous Set of Xenobiotics on Growth and Plasma Membranes of Mammalian and Fungal Cell Cultures

Cascorbi, Ingolf; Bittrich, H.; Ricklinkat, J.; Voß, Winrich; Seyfarth, Angelika; Foret, M.

doi:10.1006/eesa.1993.1043

Cited by 17 publications

(15 citation statements)

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“…The most used assay is the short-term lethality test on fish, but it is a time-consuming and costly assay and this has promoted the development of various short-term assays performed on bacteria, fungi, protozoa, or algae (Bellavere and Gorbi, 1981;Bitton, 1983;Blaise, 1991;Kupillas et al, 1991;Bragadin et al, 1992;Cascorbi et al, 1993;Belabed et al, 1994;Madoni et al, 1994;Schaefer et al, 1994). The response of the first bioassays was to provide a statement of the concentration of a substance which generated lethal effects (LD 50 ).…”

Section: Introductionmentioning

confidence: 99%

Toxicity Assessment of 16 Inorganic Environmental Pollutants by Six Bioassays

Sauvant¹,

Pépin²,

Bohatier

et al. 1997

Ecotoxicology and Environmental Safety

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Section: Introductionmentioning

confidence: 99%

Toxicity Assessment of 16 Inorganic Environmental Pollutants by Six Bioassays

Sauvant¹,

Pépin²,

Bohatier

et al. 1997

Ecotoxicology and Environmental Safety

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“…Historically, live microbes have been used as biosensors for cytotoxic environmental conditions based on changes in growth rate. Various classes of toxic compounds inhibit yeast growth in a concentration-dependent manner (8,17). Similarly, the Ames test is commonly used to assess genotoxicity by measuring Salmonella enterica serovar Typhimurium colony formation on selective medium following exposure to a chemical compound (3,11).…”

mentioning

confidence: 99%

Saccharomyces cerevisiae JEN1Promoter Activity Is Inversely Related to Concentration of Repressing Sugar

Chambers

Issaka

Palecek

2004

Appl Environ Microbiol

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When carbon sources are changed, Saccharomyces cerevisiae transcriptional patterns drastically change. To identify genes whose transcription can be used to quantitatively measure sugar concentrations, we searched genomic expression databases for a set of genes that are highly induced during the diauxic shift, and we used the promoters from these genes to drive expression of green fluorescent protein (GFP). Certain sugars, including glucose, fructose, and mannose, repress the promoter of JEN1, which encodes a lactate-pyruvate transporter, in a dose-dependent manner. Nonrepressing carbon sources include galactose, raffinose, ethanol, lactate, and glycerol. JEN1 promoter activity is a linear function of glucose concentration when organisms are grown at a steady-state glucose concentration below 1 g/liter. JEN1 promoter repression is specific to carbon source; heat or cold shock, osmotic stress, DNA damage, and nitrogen starvation do not significantly affect promoter activity. Activation of the JEN1 promoter requires the Snf1 protein kinase, but multiple regulatory elements most likely combine to provide the linear relationship between JEN1 promoter activity and sugar concentration. Thus, a JEN1 promoter-reporter system appears to provide a good living cell biosensor for the concentration of certain sugars. The JEN1 promoter also permits quantitative regulation of cellular functions not normally controlled by sugar concentrations. For example, a strain expressing FLO1 under control of the JEN1 promoter flocculates at a low glucose concentration.

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“…Saccharomyces cerevisiae has been used previously to assess toxicity through the use of an amperometric gas diffusion (oxygen) electrode, which quantifies changes in culture respiration (1,7,8,9,17). Alternatively, the effect of a compound on S. cerevisiae cultures was measured directly through inhibition of maximum growth rates (2,10). However, such toxicity assays are time-consuming and expensive in comparison to luminometry analysis.…”

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confidence: 99%

Design and Application of a Biosensor for Monitoring Toxicity of Compounds to Eukaryotes

Hollis

Killham

Glover

2000

Appl Environ Microbiol

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Here we describe an alternative approach to currently used cytotoxicity analyses through applying eukaryotic microbial biosensors. The yeast Saccharomyces cerevisiae was genetically modified to express firefly luciferase, generating a bioluminescent yeast strain. The presence of any toxic chemical that interfered with the cells' metabolism resulted in a quantitative decrease in bioluminescence. In this study, it was demonstrated that the luminescent yeast strain senses chemicals known to be toxic to eukaryotes in samples assessed as nontoxic by prokaryotic biosensors. As the cell wall and adaptive mechanisms of S. cerevisiae cells enhance stability and protect from extremes of pH, solvent exposure, and osmotic shock, these inherent properties were exploited to generate a biosensor that should detect a wide range of both organic and inorganic toxins under extreme conditions.Many luminescent bacterial biosensors have been produced which detect a wide range of pollutants while simultaneously assessing bioavailability in environmental samples. Saccharomyces cerevisiae has been used previously to assess toxicity through the use of an amperometric gas diffusion (oxygen) electrode, which quantifies changes in culture respiration (1,7,8,9,17). Alternatively, the effect of a compound on S. cerevisiae cultures was measured directly through inhibition of maximum growth rates (2, 10). However, such toxicity assays are time-consuming and expensive in comparison to luminometry analysis. Walmsley et al. (18) created an S. cerevisiae biosensor that induces green fluorescent protein expression on exposure to genotoxic agents. The luminescent biosensor designed in the present study works on a different principal (reduction in reporter gene product activity) and complements the biosensor designed by Walmsley et al. (18) by detecting a wide range of toxins, not just genotoxic agents.For the novel S. cerevisiae biosensor described here, a luminescence detection system was constructed using firefly luciferase (luc) from Photinus pyralis. The firefly luciferase light reaction relies on ATP being supplied by actively metabolizing cells. This dependence on endogenous energy supplies enables a luciferase assay system to report directly on cell health upon exposure to toxins. The luciferin substrate for P. pyralis luciferase is an amphipathic molecule with a charged carboxyl group at physiological pH. This prevents easy passage of luciferin across cell membranes, leading to problems during the exogenous addition for in vivo assays. This was overcome through the development of a novel assay system where the biosensor preparation was acidified after exposure to the toxicant and before luminescence quantification. MATERIALS AND METHODSStrains, media, and chemicals. S. cerevisiae strain W303-1B 15 trp1-1 can1-100 ade2-1 ura3-1) (16) was the host for the chromosomal luciferase-expressing construct. Chromosomal insertion was carried out using a published method (5). S. cerevisiae was grown in synthetic complete medium (15)

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Effects of a Heterogenous Set of Xenobiotics on Growth and Plasma Membranes of Mammalian and Fungal Cell Cultures

Cited by 17 publications

References 0 publications

Toxicity Assessment of 16 Inorganic Environmental Pollutants by Six Bioassays

Toxicity Assessment of 16 Inorganic Environmental Pollutants by Six Bioassays

Saccharomyces cerevisiae JEN1Promoter Activity Is Inversely Related to Concentration of Repressing Sugar

Design and Application of a Biosensor for Monitoring Toxicity of Compounds to Eukaryotes

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