Mutagenic potency of exocyclic DNA adducts: marked differences between Escherichia coli and simian kidney cells.
A single-stranded shuttle vector containing a single 3,N4-etheno-2'-deoxycytidine (epsilon dC) or 1,N2-(1,3-propano)-2'- deoxyguanosine (PdG) DNA adduct was used to investigate translesional DNA synthesis in Escherichia coli and simian kidney (COS) cells. The presence of either exocyclic adduct was associated with reduced numbers of transformants. In E. coli, this inhibitory effect could be overcome partially by irradiating cells with UV light before transformation. Translesional synthesis past both exocyclic lesions was accompanied by targeted mutations. For PdG, the primary mutagenic events observed in both hosts were PdG-->T transversions; in preirradiated E. coli, PdG-->A transitions were also observed. The targeted mutation frequency for single-stranded DNA that contained PdG was 100% in nonirradiated E. coli, 68% in preirradiated cells, and 8% in COS cells. In contrast, the targeted mutation frequency for single-stranded DNA that contained epsilon dC was 2% in nonirradiated E. coli, 32% in preirradiated cells, and 81% in COS cells. The primary mutations generated by epsilon dC in both E. coli and COS cells were epsilon dC-->A and epsilon dC-->T base substitutions. These observations appear to reflect the variable specificity of DNA replication complexes in incorporating bases opposite certain adducts. We conclude that DNA synthesis past the same DNA adduct can have strikingly different consequences in bacteria and mammalian cells, underscoring the importance of establishing the intrinsic mutagenic potential of DNA adducts in mammalian cells.
Moriya
M
M
Department of Pharmacological Sciences, State University of New York, Stony Brook 11794-8651.
Zhang
W
W
Johnson
F
F
Grollman
A P
AP
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002461:Cell Line, Transformed; D002522:Chlorocebus aethiops; D018736:DNA Adducts / Q000494:pharmacology*; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000187:drug effects; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics / Q000528:radiation effects; D007668:Kidney; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009152:Mutagenicity Tests; D009153:Mutagens / Q000494:pharmacology*; D013539:Simian virus 40 / Q000235:genetics; D014466:Ultraviolet Rays
C090855:1,N(2)-(1,3-propano)-2'-deoxyguanosine; D018736:DNA Adducts; D004277:DNA, Single-Stranded; D009153:Mutagens; D003841:Deoxycytidine; C032222:3,N(4)-ethenodeoxycytidine; D003849:Deoxyguanosine