Detecting AhR ligands in sediments using bioluminescent reporter yeast.

Sediments polluted with high concentrations of persistent organic pollutants, many of which are ligands of the aryl hydrocarbon receptor (AhR), are currently of concern around the industrialized world. Bioassays that can detect the presence of AhR ligands in environmental samples offer a relatively rapid and cost-effective means of prioritizing samples before more elaborate, laborious, and costly chemical analyses are applied. This paper presents a new bioluminescent yeast assay based on transcriptional activation of AhR. Its applicability for determining AhR ligands in complex environmental samples was demonstrated by analyzing a set of sediment samples from the River Kymi, Finland. The results from the assay are shown to be consistent with those from both a chemical analysis and an H4IIE-luc bioassay. The yeast assay procedure is simple and can be performed within 1 day. The yeasts grow rapidly, are easy to handle, and do not require continuous cell culturing. Moreover, the robustness of the yeast allows the application of the test to crude extracts or even sediment suspensions. The yeast assay described in this paper can be useful in screening and prioritization of samples prior to chemical analysis. Moreover, the strain can be used in the construction of fibre-optic biosensors.

[1]  Steven Ripp,et al.  Expression of the Photorhabdus luminescens lux genes (luxA, B, C, D, and E) in Saccharomyces cerevisiae. , 2003, FEMS yeast research.

[2]  Uwe Spohn,et al.  Photodiode-based chemiluminometric biosensors for hydrogen peroxide and L-lysine , 1994 .

[3]  S. Safe,et al.  Polychlorinated biphenyls (PCBs), dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and related compounds: environmental and mechanistic considerations which support the development of toxic equivalency factors (TEFs). , 1990, Critical reviews in toxicology.

[4]  J. Vondráček,et al.  Aryl hydrocarbon receptor-mediated activity of mutagenic polycyclic aromatic hydrocarbons determined using in vitro reporter gene assay. , 2001, Mutation research.

[5]  M. Karp,et al.  Reporter genes lucFF, luxCDABE, gfp, and dsred have different characteristics in whole-cell bacterial sensors. , 2002, Analytical biochemistry.

[6]  C. Miller.  A human aryl hydrocarbon receptor signaling pathway constructed in yeast displays additive responses to ligand mixtures. , 1999, Toxicology and applied pharmacology.

[7]  Eliora Z Ron,et al.  Biosensing environmental pollution. , 2007, Current opinion in biotechnology.

[8]  O. Hankinson.  The aryl hydrocarbon receptor complex. , 1995, Annual review of pharmacology and toxicology.

[9]  Alessandro Pandini,et al.  Ligand binding and activation of the Ah receptor. , 2002, Chemico-biological interactions.

[10]  J. Giesy,et al.  Chemical-activated luciferase gene expression (CALUX): a novel in vitro bioassay for Ah receptor active compounds in sediments and pore water. , 1996, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[11]  Michael J. Paulus,et al.  Bioluminescent-bioreporter integrated circuits form novel whole-cell biosensors , 1998 .

[12]  Marko Virta,et al.  Detection of bioavailable heavy metals in EILATox‐Oregon samples using whole‐cell luminescent bacterial sensors in suspension or immobilized onto fibre‐optic tips , 2004, Journal of applied toxicology : JAT.

[13]  P. Shankar,et al.  A review of fiber-optic biosensors , 2007 .

[14]  J. Beasley,et al.  Archives of environmental contamination and toxicology , 1978 .

[15]  M. Denison,et al.  Activation of the aryl hydrocarbon receptor by structurally diverse exogenous and endogenous chemicals. , 2003, Annual review of pharmacology and toxicology.

[16]  Bruce D Hammock,et al.  Development of a green fluorescent protein-based cell bioassay for the rapid and inexpensive detection and characterization of ah receptor agonists. , 2002, Toxicological sciences : an official journal of the Society of Toxicology.

[17]  Paul D. Jones,et al.  Cell bioassays for detection of aryl hydrocarbon (AhR) and estrogen receptor (ER) mediated activity in environmental samples. , 2002, Marine pollution bulletin.

[18]  H. Mussalo-Rauhamaa,et al.  Polychlorinated diphenyl ethers, dibenzo-P-dioxins and dibenzofurans in Finnish human tissues compared to environmental samples. , 1995, Chemosphere.

[19]  A. Baudin-Baillieu,et al.  Construction of a Yeast Strain Deleted for the TRP1 Promoter and Coding Region that Enhances the Efficiency of the Polymerase Chain Reaction‐Disruption Method , 1997, Yeast.

[20]  L. Poellinger,et al.  Heat shock protein hsp90 regulates dioxin receptor function in vivo. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[21]  H. Sheppard,et al.  Transcriptional activation by estrogen receptor (ERalpha) and steroid receptor coactivator (SRC1) involves distinct mechanisms in yeast and mammalian cells. , 2003, Journal of molecular endocrinology.

[22]  A. Roda,et al.  Bio- and chemiluminescence in bioanalysis , 2000, Fresenius' journal of analytical chemistry.

[23]  O. Hankinson,et al.  Recruitment of the NCoA/SRC-1/p160 Family of Transcriptional Coactivators by the Aryl Hydrocarbon Receptor/Aryl Hydrocarbon Receptor Nuclear Translocator Complex , 2002, Molecular and Cellular Biology.

[24]  J. Giesy,et al.  H4IIE rat hepatoma cell bioassay-derived 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents in colonial fish-eating waterbird eggs from the Great Lakes , 1991, Archives of environmental contamination and toxicology.

[25]  C. Bradfield,et al.  Ah receptor signaling pathways. , 1996, Annual review of cell and developmental biology.

[26]  J. D. Winefordner,et al.  Limit of detection. A closer look at the IUPAC definition , 1983 .

[27]  L. Mendonça-Hagler,et al.  Trends in biotechnology and biosafety in Brazil. , 2008, Environmental biosafety research.

[28]  Elisa Michelini,et al.  Bioluminescent yeast assays for detecting estrogenic and androgenic activity in different matrices. , 2005, Chemosphere.

[29]  L. Hyman,et al.  Assessment of aryl hydrocarbon receptor complex interactions using pBEVY plasmids: expressionvectors with bi-directional promoters for use in Saccharomyces cerevisiae. , 1998, Nucleic acids research.

[30]  C. Bradfield,et al.  The 90-kDa heat shock protein is essential for Ah receptor signaling in a yeast expression system. , 1994, The Journal of biological chemistry.

[31]  J. Giesy,et al.  Relative Potencies of Individual Polychlorinated Naphthalenes to Induce Dioxin-Like Responses in Fish and Mammalian In Vitro Bioassays , 2000, Archives of environmental contamination and toxicology.

[32]  A. Oikari,et al.  Occurrence of retene and resin acids in sediments and fish bile from a lake receiving pulp and paper mill effluents , 1999 .

[33]  Matti Verta,et al.  Spatial distibution and temporal accumulation of polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls in the Gulf of Finland. , 2002, Environmental science & technology.

[34]  Y. Shimizu,et al.  Characterization of polycyclic aromatic hydrocarbons (PAHs) in different size fractions in deposited road particles (DRPs) from Lake Biwa area, Japan. , 2005, Environmental science & technology.

[35]  M. Tamminen,et al.  Quantification of ecotoxicological tests based on bioluminescence using Polaroid film. , 2007, Chemosphere.

[36]  J. Giesy,et al.  Relative potencies of individual polycyclic aromatic hydrocarbons to induce dioxinlike and estrogenic responses in three cell lines , 2002, Environmental toxicology.

[37]  H. Shiraishi,et al.  Luminescent yeast cells entrapped in hydrogels for estrogenic endocrine disrupting chemical biodetection. , 2006, Biosensors & bioelectronics.

[38]  L. Birnbaum,et al.  Chemically activated luciferase gene expression (CALUX) cell bioassay analysis for the estimation of dioxin-like activity: critical parameters of the CALUX procedure that impact assay results. , 2005, Environmental science & technology.

[39]  M. Karp,et al.  One‐step measurement of firefly luciferase activity in yeast , 2003, Yeast.

[40]  J. Giesy,et al.  Species-specific recombinant cell lines as bioassay systems for the detection of 2,3,7,8-tetrachlorodibenzo-p-dioxin-like chemicals. , 1996, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[41]  John P. Giesy,et al.  Derivation and application of relative potency estimates based on in vitro bioassay results , 2000 .