METALLOTHIONEIN PROTECTS DNA FROM OXIDATIVE DAMAGE

Metallothionein (MT) is a potent hydroxyl radical scavenger but its antioxidant properties in vivo have not been defined. Most of the recent results indicate that it does not afford protection to cells against the lethal action of oxidative stress. However, the possibility that MT confers protection against oxidative damage to a specific cellular target, such as DNA, had not been considered. We compared V79 Chinese hamster cells enriched in and depleted of MT in terms of DNA-strand scission. Zinc induces an increase in MT content of V79 Chinese hamster cells, without concomitant increase in the GSH level. These induced cells are more resistant to the production of DNA-strand scission by H2O2 than the parental cells. Conversely, cells rendered partially deprived of MT, by transfection with a plasmid vector in which the MT-I cDNA is antisense oriented in relation to a simian virus 40 promoter, became more susceptible to the DNA-damaging action of H2O2. The transfected cells did not exhibit alterations of GSH, superoxide dismutase- and H2O2-destroying enzymes. Indirect immunofluorescence indicated that most of the MT was concentrated in the cell nucleus. Neither overexpression nor lower expression of MT resulted in differential resistance to the killing action of H2O2. However, the combined high nuclear concentration of MT and its excellent hydroxyl scavenger properties confer protection to DNA from hydroxyl radical attack.