INDUCED SYNTHESIS OF O-6-METHYLGUANINE-DNA METHYLTRANSFERASE IN RAT HEPATOMA-CELLS EXPOSED TO DNA-DAMAGING AGENTS

When the rat hepatoma cell line H4IIE was treated with DNA-damaging agents such as N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), ultraviolet light and gamma-rays, the O6-methylguanine-DNA methyltransferase activity increased 2 to 3 times over the level seen in non-treated cells. SDS/polyacrylamide gel electrophoresis followed by fluorography revealed that a single species of methyltransferase protein with a molecular weight of 25,500 was present in both non-treated and treated cells. Northern blot analysis using a cloned rat cDNA as a probe revealed that the enzyme activity increased because transcription of the gene was enhanced. The level of enzyme activity increased within 48 h after UV irradiation and remained at a higher level for 150 h. Following UV irradiation, the cells become more resistant than the normal cells to MNNG.