[en] The Chinese hamster ovary cell line CHO-T510 contains a single copy of a stably integrated retroviral vector with a selectable marker, the E. coli xanthine-guanine phosphoribosyl transferase (gpt) gene. Previous studies on the CHO-T510 line showed that, in comparison with other genetic loci, the gpt locus was hypersensitive to mutation induction by ionizing radiation. Southern blot analyses of a set of 20-26 gpt^- mutant lines, isolated as either spontaneous, gamma-induced, or alpha-radiation-induced mutants, indicated that 86-95% of these were complete vector deletions. The integrated gpt vector was localized by in situ hybridization to the q arm of chromosome 5 in close proximity to the interstitial ttelomeresequences near the pericentric region of this chromosome. One to three kilobases of sequences adjacent to the gpt integration site were clones and analyzed. Both the right and left integration sites contain sequences that hybridize to a pantelomere probe, suggesting that the vector has acquired telomeric repeats at its ends. The radio-sensitivity of the gpt locus may be due to these telomere repeats, as interstitial telomeres have been reported to be radiation-sensitive fragile sites. The gpt locus in the T510 line affords a unique resource to test this hypothesis

[en] We have been characterizing a Chinese hamster ovary cell line, CHO-K1 10T5, into which a gpt-containing retroviral shuttle vector has been stably integrated. This normally stable locus in CHO-K1 10T5 cells is very sensitive to deletion mutation following ionizing radiation exposure and shows an LET response with an RBE of 3 for α particles. Almost all of the gpt mutants are total gene deletions. The gpt gene has been localized to chromosome 5q15, within 100-1000 kb of a large region of interstitial telomere repeats. In addition, sequences showing homology to telomere repeat sequences have been identified at the integration site both 5' and 3' to the gpt gene locus. The integration site has been cloned and is presently being sequenced. Preliminary data suggest that there is a hotspot for breakage and/or recombination 5' of the integration site. We hypothesize that (1) the gpt vector has integrated into a region containing telomere repeat sequences, (2) this region of the genome is a radiation-sensitive fragile site, and (3) the radiation sensitivity of this site is due to the telomere sequences, which act by either serving as a site for further telomerase action and chromosome terminilization, or by providing repeat structures to facilitate radiation-induced recombination