Monitoring of endocrine disruptors in surface waters by the yeast recombinant assay

Endocrine disruptors exert physiological effects at very low concentrations. Surface waters present often a mixture of high concentrations of low-potency disruptors and low amounts of very powerful ones, making their chemical analysis complicated and expensive. We developed a recombinant yeast assay (RYA) for estrogenic compounds using 96-well microtiter plates. This assay is based on three yeast strains, transformed with self-propagating plasmids. One strain contains an expression plasmid for the human estrogen hormone receptor and an appropriate reporter; it detects estrogenic and antiestrogenic activities. The two other yeast strains, one expressing the human progesterone receptor and a second based on the yeast activator Gal4p, served to analyze the nature of antiestrogenic activities. We applied this technique to water samples from two tributaries on the Llobregat river (NE Spain) as well as from four sewage treatment plants discharging on them. Our results indicate that the efficiency of sewage treatment plants for eliminating estrogenic compounds varied notably, being in at least one case completely inefficient. We also observed a prevalence of an inhibitory activity all through the two rivers; this inhibition was hormone specific. These results were consistent to previously obtained chemical analyses of the same samples, although chemical and in vivo analyses showed rather different levels of sensitivity for some compounds. Our findings demonstrate the utility of the yeast recombinant assay for analyzing complex natural samples; at the same time, they stress the necessity of a panel of different yeast systems to adequately describe endocrine-disruptor activities.

[1]  B. O’Malley,et al.  Ligand-dependent and -independent function of the transactivation regions of the human estrogen receptor in yeast. , 1992, Molecular endocrinology.

[2]  L. Guarente,et al.  Vectors for expression of cloned genes in yeast: regulation, overproduction, and underproduction. , 1991, Methods in enzymology.

[3]  D. Barceló,et al.  Identification of polar, ionic, and highly water soluble organic pollutants in untreated industrial wastewaters , 1999 .

[4]  L. Jin,et al.  Evaluation of clinical and environmental anti-estrogens with human estrogen receptor expressed in Saccharomyces cerevisiae: a novel role for ABC-cassette transporters in mediating anti-estrogenic activity. , 1997, Biochemical and Biophysical Research Communications - BBRC.

[5]  C. Portier,et al.  Evaluation of chemicals with endocrine modulating activity in a yeast-based steroid hormone receptor gene transcription assay. , 1997, Toxicology and applied pharmacology.

[6]  Edwin J. Routledge,et al.  Identification of Estrogenic Chemicals in STW Effluent. 2. In Vivo Responses in Trout and Roach , 1998 .

[7]  Hong Wang,et al.  Activation of the human estrogen receptor by the antiestrogens ICI 182,780 and tamoxifen in yeast genetic systems: implications for their mechanism of action. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[8]  N. Coldham,et al.  Evaluation of a recombinant yeast cell estrogen screening assay. , 1997, Environmental health perspectives.

[9]  F. Sherman Getting started with yeast. , 1991, Methods in enzymology.

[10]  L. Guarente,et al.  Heme regulates transcription of the CYC1 gene of S. cerevisiae via an upstream activation site , 1983, Cell.

[11]  D. Barceló,et al.  Determination of steroid sex hormones and related synthetic compounds considered as endocrine disrupters in water by liquid chromatography-diode array detection-mass spectrometry. , 2000, Journal of chromatography. A.

[12]  Miguel Beato,et al.  Steroid hormone receptors: Many Actors in search of a plot , 1995, Cell.

[13]  W. House,et al.  Environmental fate of nonylphenol ethoxylates: Differential adsorption of homologs to components of river sediment , 2000 .

[14]  W. Verstraete,et al.  Nonylphenol and Estrogenic Activity in Aquatic Environmental Samples , 1999 .

[15]  T. Butt,et al.  Yeast hormone response element assays detect and characterize GRIP1 coactivator-dependent activation of transcription by thyroid and retinoid nuclear receptors. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[16]  J. Cravedi,et al.  Two complementary bioassays for screening the estrogenic potency of xenobiotics: recombinant yeast for trout estrogen receptor and trout hepatocyte cultures. , 1997, Journal of molecular endocrinology.

[17]  K. Umesono,et al.  The nuclear receptor superfamily: The second decade , 1995, Cell.

[18]  Mark Johnston,et al.  5 Regulation of Carbon and Phosphate Utilization , 1992 .

[19]  Todd F. Wheeler,et al.  Mass Spectral Characterization of p-Nonylphenol Isomers Using High-Resolution Capillary GC—MS , 1997 .

[20]  K. Schramm,et al.  Applicability of a yeast oestrogen screen for the detection of oestrogen-like activities in environmental samples. , 1999, Chemosphere.

[21]  J. McLachlan,et al.  The anti-estrogenic activity of selected polynuclear aromatic hydrocarbons in yeast expressing human estrogen receptor. , 1996, Biochemical and biophysical research communications.

[22]  S. Harrison,et al.  DNA sequence preferences of GAL4 and PPR1: how a subset of Zn2 Cys6 binuclear cluster proteins recognizes DNA , 1996, Molecular and cellular biology.

[23]  B. Piña,et al.  Structural and functional heterogeneity of Rap1p complexes with telomeric and UASrpg-like DNA sequences. , 1998, Journal of molecular biology.

[24]  D. Barceló,et al.  Estrogenicity determination in sewage treatment plants and surface waters from the Catalonian area (NE Spain) , 2000 .

[25]  J. McLachlan,et al.  The estrogenic and antiestrogenic activities of phytochemicals with the human estrogen receptor expressed in yeast , 1997, Steroids.

[26]  C. Sonnenschein,et al.  p-Nonyl-phenol: an estrogenic xenobiotic released from "modified" polystyrene. , 1991, Environmental health perspectives.

[27]  John P. Sumpter,et al.  Detergent components in sewage effluent are weakly oestrogenic to fish: An in vitro study using rainbow trout (Oncorhynchus mykiss) hepatocytes , 1993 .