A new Salmonella typhimurium NM5004 strain expressing rat glutathione S-transferase 5–5: use in detection of genotoxicity of dihaloalkanes using an SOS/umu test system

Yoshimitsu Oda, Hiroshi Yamazaki, Ricarda Thier, Brian Ketterer, F.Peter Guengerich, Tsutomu Shimada
1996 Carcinogenesis  
The Escherichia coli mu operon was subcloned into a pKK233-2 vector containing rat glutathione S-transferase (GST) 5-5 cDNA and the plasmid thus obtained was introduced into Salmonella typhimurium TA1535. The newly developed strain S.typhimurium NM5004, was found to have 52-fold greater GST activity than the original umu strain S.typhimurium TA1535/pSK1002. We compared sensitivities of these two tester strains, NM5004 and TA1535/ pSK1002, for induction of umuC gene expression with several
more » ... alkanes which are activated or inactivated by GST 5-5 activity. The induction of umuC gene expression by these chemicals was monitored by measuring the cellular p*-galactosidase activity produced by umuC'lacZ fusion gene in these two tester strains. Ethylene dibromide, 1bromo-2-chloroethane, 1,2-dichloroethane, and methylene dichloride induced umuC gene expression more strongly in the NM5004 strain than the original strain. 4-Nltroquinoline 1-oxide and iV-methyl-yV'-nitro-iV-nitrosoguanidine were found to induce umuC gene expression to similar extents in both strains. In the case of 1-nitropyrene and 2nitrofluorene, however, NM5004 strain showed weaker umuC gene expression responses than the original TA1535/ pSK1002 strain. l,2-Epoxy-3-(4'-nitrophenoxy)propane, a known substrate for GST 5-5, was found to inhibit umuC induction caused by l-bromo-2-chloroethane. These results indicate that this new tester NM5004 strain expressing a mammalian GST theta class enzyme may be useful for studies of environmental chemicals proposed to be activated or inactivated by GST activity.
doi:10.1093/carcin/17.2.297 pmid:8625454 fatcat:xnzbf3ixpbdgzdwa5zry2jrp6y