Effects of O (6)-alkylguanine-DNA alkyltransferase deficiency in Escherichia coli as the host for the detection of mutations in lacI transgenic mice.
Transgenic mice are widely used to detect gene mutations in vivo induced by a variety of chemicals. It is known, however, that no mutagenicity of methyl methanesulfonate (MMS) is detected in epididymal sperm in various transgenic mice assays, although MMS induces the dominant lethal and specific locus mutations in male mice. To investigate the issue of whether unrepaired lesions in DNA of mature sperm can be transformed into mutations during replication of the lambda phage in Escherichia coli cells, we developed an E. coli strain YG5152, which is a derivative of strain SCS-8 but is deficient in the genes encoding O (6)-alkylguanine-DNA alkyltransferases. When lambda LIZalpha phages were treated with MMS or N-ethyl-N-nitrosourea (ENU) in vitro and infected to the E. coli strains, the mutant frequencies of lacI were markedly higher in strain YG5152 than in strain SCS-8. When Big Blue(trade mark) mice were treated with MMS (160 mg/kg) or ENU (125 or 250 mg/kg) and the phages rescued from mature sperm were infected to the strains, the mutation frequency (MF) of phages from ENU-treated mice at a dose of 250 mg/kg in strain YG5152 was about two times higher than that in strain SCS-8. However, no increase in the MF was observed in the MMS-treated mice even in strain YG5152. These results suggest that, although strain YG5152 efficiently detects ex vivo mutations caused by mutagenic alkyl adducts formed by MMS in lambda phage DNA, no detectable levels of mutagenic methyl adducts are present in mature sperm of MMS-treated mice. Possible reasons for this lack of mutagenicity of MMS in mature sperm using transgenic mice assays are discussed.
Sui
H
H
Hatano Research Institute, Food and Drug Safety Center, Ochiai, Hadano-shi, Kanagawa, Japan. sui@ma3.justnet.ne.jp
Suzuki
M
M
Yamada
M
M
Hara
T
T
Kawakami
K
K
Shibuya
T
T
Nohmi
T
T
Sofuni
T
T
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D010582:Bacteriophage lambda / Q000187:drug effects / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins; D005038:Ethylnitrosourea / Q000494:pharmacology; D056968:Lac Repressors; D008297:Male; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D051379:Mice; D008822:Mice, Transgenic; D009153:Mutagens / Q000494:pharmacology; D009154:Mutation; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics*; D013094:Spermatozoa / Q000187:drug effects
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D056968:Lac Repressors; C539064:LacI protein, E coli; D009153:Mutagens; D012097:Repressor Proteins; D008741:Methyl Methanesulfonate; D019853:O(6)-Methylguanine-DNA Methyltransferase; D005038:Ethylnitrosourea