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Sensitive detection of chemical‐induced genotoxicity by the Cypridina secretory luciferase reporter assay, using DNA repair‐deficient strains of Saccharomyces cerevisiae
Author(s) -
Ochi Yukari,
Sugawara Harumi,
Iwami Mio,
Tanaka Megumi,
Eki Toshihiko
Publication year - 2011
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1837
Subject(s) - genotoxicity , saccharomyces cerevisiae , luciferase , biology , reporter gene , plasmid , dna damage , yeast , dna , microbiology and biotechnology , mutant , dna repair , biochemistry , gene , chemistry , gene expression , transfection , toxicity , organic chemistry
Yeast‐based reporter assays are useful for detecting various genotoxic chemicals. We established a genotoxicity assay using recombinant strains of Saccharomyces cerevisiae , each containing a reporter plasmid with the secretory luciferase gene from Cypridina noctiluca , driven by a DNA damage‐responsive promoter of the yeast RNR3 gene. This system detected the genotoxicity of methyl methanesulphonate (MMS) as sensitively as conventional yeast‐based reporter assays, using the β‐galactosidase gene in a concentration‐dependent manner; it also detects four other genotoxic chemicals, allowing us to monitor DNA damage easily by skipping the cell extraction process for the assay. We examined Cypridina luciferase levels induced by MMS and three antitumour agents using a set of BY4741‐derived deletion mutants, each defective in a DNA repair pathway or DNA damage checkpoint. Luciferase activities were particularly enhanced in mutant strains with mms2Δ and mag1Δ by exposure to MMS, rad59Δ and mlh1Δ to camptothecin and mms2Δ and mlh1Δ to mitomycin C, respectively, compared with their parent strains. Enhanced reporter activities were also found in some DNA repair mutants with cisplatin. These observations suggest that this Cypridina secretory luciferase reporter assay using yeast DNA repair mutants offers convenient and sensitive detection of the potential genotoxicity of numerous compounds, including antitumour drugs and studying the mechanisms of DNA damage response in yeast. Copyright © 2011 John Wiley & Sons, Ltd.