Chromosome terminal deletion formation in Chinese hamster ovary cells

To investigate the fate of unrejoined DNA double-strand breaks, the frequency of 60Co gamma-ray- and restriction-enzyme-induced terminal chromosome deletions, a marker of unrejoined breaks, was determined in CHO-K1 and in xrs-5 cells. The xrs-5 cell is a DNA double-strand break repair-deficient derivative of CHO-K1. Terminal deletion frequency was small in both CHO-K1 and xrs-5 cells when cells were irradiated or treated with restriction enzyme while in the G1 phase of the cell cycle. In contrast, previous studies have shown that treatment of cells in G2 leads to large deletion frequencies, especially in xrs-5 cells. Cell cycle analyses show large G2 blocks in irradiated xrs-5 cells with only partial recovery over a 24-96-h period. These results suggest that most CHO cells with unrejoined breaks are blocked in G2 and, therefore, do not contribute to chromosome mutation frequencies. The small frequencies of terminal deletions that are found in these cells may reflect either an inefficiency in the G2 checkpoint mechanism or, perhaps, a modification of broken ends that allows passage through G2.