Electrophoretically detectable mutations induced in CHO cells by varying doses of ultraviolet radiation

Chinese hamster ovary (CHO) cells were subjected to ultraviolet radiation (UV) at doses resulting in 100% (no irradiation), 50-30%, 20-10% and approximately 1% survival. 2 divisions after UV exposure surviving cells were cloned and clones expanded for electrophoretic analysis of the products of approximately 40 enzyme loci. 4 different classes of variants (electrophoretic shifts, nulls, enzyme re-expression and enzyme modification) were detected in 29 of 1329 clones analyzed and proven mutants by subclone analysis. The frequency of mutants in the irradiated groups (28/38391 loci screened or 7.3 X 10(-4) was significantly higher than controls. The frequency of shift mutants at 10-20% survival was higher than shifts at 30-50% survival and was significantly higher than shifts at approximately 1% survival. The frequency of nulls increased with dose. 12 of the 28 mutants obtained in the irradiated groups were at only 3 of the mean 41 loci screened/clone. These results indicated that shift mutants could be detected more efficiently than nulls at lower dose and that loci varied widely with respect to their susceptibility to UV mutagenesis. Multiple null mutants at 2 loci, isocitrate dehydrogenase 2 and hexokinase 2, indicated they may be hemizygous in CHO cells.