Expression in mammalian cells of the Escherichia coli O6 alkylguanine-DNA-alkyltransferase gene ogt reduces the toxicity of alkylnitrosoureas.
V79 Chinese hamster cells expressing either the O6-alkylguanine-DNA-alkyltransferase (ATase) encoded by the E. coli ogt gene or a truncated version of the E. coli ada gene have been exposed to various alkylnitrosoureas to investigate the contribution of ATase repairable lesions to the toxicity of these compounds. Both ATases are able to repair O6-alkylguanine (O6-AlkG) and O4-alkylthymine (O4-AlkT) but the ogt ATase is more efficient in the repair of O4-methylthymine (O4-MeT) and higher alkyl derivatives of O6-AlkG than is the ada ATase. Expression of the ogt ATase provided greater protection against the toxic effects of the alkylating agents then the ada ATase particularly with N-ethyl-N-nitrosourea (ENU) and N-butyl-N-nitrosourea (BNU) to which the ada ATase expressing cells were as sensitive as parent vector transfected cells. Although ogt was expressed at slightly higher levels than the truncated ada in the transfected cells, this could not account for the differential protection observed. For-N-methyl-N-nitrosourea (MNU) the increased protection in ogt-transfected cells is consistent with O4-MeT acting as a toxic lesion. For the longer chain alkylating agents and chloroethylating agents, the protection afforded by the ogt protein may be a consequence of the more efficient repair of O6-AlkG, O4-AlkT or both of these lesions in comparison with the ada-encoded ATase.
Harris
L C
LC
CRC Department of Carcinogenesis, Paterson Institute for Cancer Research, Christie Hospital, Manchester, UK.
Margison
G P
GP
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002470:Cell Survival / Q000502:physiology; D002999:Clone Cells; D006224:Cricetinae; D004351:Drug Resistance; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D015870:Gene Expression / Q000235:genetics*; D006147:Guanine / Q000031:analogs & derivatives / Q000502:physiology; D008168:Lung / Q000166:cytology; D008780:Methyltransferases / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009607:Nitrosourea Compounds / Q000633:toxicity*; D019853:O(6)-Methylguanine-DNA Methyltransferase; D013941:Thymine / Q000031:analogs & derivatives / Q000502:physiology; D014162:Transfection
D001426:Bacterial Proteins; D009607:Nitrosourea Compounds; C021223:O-4-methylthymine; D006147:Guanine; C008449:O-(6)-methylguanine; D008780:Methyltransferases; D019853:O(6)-Methylguanine-DNA Methyltransferase; D013941:Thymine