title: Genotoxicity of chemical compounds identification and assessment by yeast cells transformed with GFP reporter constructs regulated by the PLM2 or DIN7 promoter.
abstract: Yeast cells transformed with high-copy number plasmids comprising a green fluorescent protein (GFP)-encoding gene optimized for yeast under the control of the new DIN7 or PLM2 and the established RNR2 and RAD54 promoters were used to assess the genotoxic potential of chemical compounds. The activity of potential DNA-damaging agents was investigated by genotoxicity assays and by OxoPlate assay in the presence of various test compounds. The fluorescence signal generated by GFP in response to DNA damage was related to the different concentrations of analytes and the analyte-dependent GFP synthesis. The use of distinct DNA damage-inducible promoters presents alternative genotoxicity testing strategies by selective induction of promoters in response to DNA damage. The new DIN7 and PLM2 systems show higher sensitivity than the RNR2 and RAD54 systems in detecting 4-nitroquinoline-N-oxide and actinomycin D. Both DIN7 and PLM2 systems are able to detect camptothecin while RNR2 and RAD54 systems are not. Automated laboratory systems with assay performance on 384-well microplates provide for cost-effective high-throughput screening of DNA-damaging agents, reducing compound consumption to about 53% as compared with existing eukaryotic genotoxicity bioassays.
authors.affiliation: Institute of Biotechnology, Vietnam Academy of Science and Technology (VAST), Hanoi, Vietnam.
authors.forename: Van Ngoc
authors.initials: VN
authors.lastname: Bui
authors.affiliation: Thai Nguyen University of Sciences, Thai Nguyen University, Thai Nguyen, Hanoi, Vietnam.
authors.forename: Thi Thu Huyen
authors.initials: TT
authors.lastname: Nguyen
authors.affiliation: Institute of Research and Development, UMR ECOSYM, Montpellier, France.
authors.forename: Yvan
authors.initials: Y
authors.lastname: Bettarel
authors.affiliation: Institute of Biotechnology, Vietnam Academy of Science and Technology (VAST), Hanoi, Vietnam.
authors.forename: Thi Hoai Thu
authors.initials: TH
authors.lastname: Nguyen
authors.affiliation: Institute of Biotechnology, Vietnam Academy of Science and Technology (VAST), Hanoi, Vietnam.
authors.forename: Thuy Linh
authors.initials: TL
authors.lastname: Pham
authors.affiliation: Institute of Biotechnology, Vietnam Academy of Science and Technology (VAST), Hanoi, Vietnam.
authors.forename: Thi Yen
authors.initials: TY
authors.lastname: Hoang
authors.affiliation: Thai Nguyen University of Sciences, Thai Nguyen University, Thai Nguyen, Hanoi, Vietnam.
authors.forename: Vu Thanh Thanh
authors.initials: VT
authors.lastname: Nguyen
authors.affiliation: Institute of Biotechnology, Vietnam Academy of Science and Technology (VAST), Hanoi, Vietnam.
authors.forename: Ngoc Minh
authors.initials: NM
authors.lastname: Nghiem
authors.affiliation: Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Heidelberg, Germany wolfl@uni-hd.de.
authors.forename: Stefan
authors.initials: S
authors.lastname: Wölfl
mesh_terms: D004265:DNA Helicases / Q000235:genetics; D045643:DNA Repair Enzymes / Q000235:genetics; D005090:Exodeoxyribonucleases / Q000235:genetics*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D009152:Mutagenicity Tests; D009153:Mutagens / Q000633:toxicity*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012264:Ribonucleotide Reductases / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*
chemical_list: D009153:Mutagens; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; C570014:RNR2 protein, S cerevisiae; D012264:Ribonucleotide Reductases; C105071:DIN7 protein, S cerevisiae; D005090:Exodeoxyribonucleases; C075468:RAD54 protein, S cerevisiae; D004265:DNA Helicases; D045643:DNA Repair Enzymes
keywords: DNA damage-inducible promoters; DNA-damaging agents; GFP; biosensor; genotoxicity