Summary of complementation groups of UV-sensitive CHO cell mutants isolated by large-scale screening

Abstract

A summary is given for the lineage and complementation group assignments of 153 UV-sensitive mutants of the CHO AA8 cell line. The distribution of mutants among six complementation groups was highly non-random, with the great majority of the isolates belonging to groups 1 and 2. This asymmetry is consistent with the known hemizygosity of these two linked loci in CHO cells. The relative numbers of mutants induced in group 2 was found to depend greatly on the type of mutagen used. Mutagenesis with UV radiation, ethyl methanesulfonate (EMS), N-methyl-N' -nitro-N-nitroso-guanidine and 7-bromomethylbenz[a]anthracene produced high frequencies of group 2 mutants. In contrast, ICR170 and ICR191, which are thought to produce mostly frameshift mutations, yielded very few mutants in group 2. These results are of particular importance in light of the recent finding that the human ERCC2 gene, which corrects group 2 mutants, has very strong homology with the yeast gene RAD3. RAD3 is an essential gene for viability in yeast, and the low recovery of group 2 mutants using the frameshift agents strongly suggests that frameshift mutations tend to be lethal in the hamster ERCC2 locus. Several mutagen-sensitive double mutants were isolated in two-step selections from EMS-, mitomycin C- or UV-sensitive parental cells, including the line UVU1, the first mammalian line with two mutations that affect UV sensitivity. The first mutation inactivated excision repair, and the second mutation appears to have affected some other recovery process. UVU1 should be useful for studying recovery processes that are separate from nucleotide excision repair.