Mutagen-induced recombination between stably integrated neo gene fragments in CHO and EM9 cells.
The ability of mutagenic agents to induce homologous recombination was studied in a 'normal' Chinese hamster ovary (CHO) cell clone (CHO:5) and a cell clone (EM9:2) derived from the presumptive DNA repair-deficient mutant cell line EM9, which has a high spontaneous sister-chromatid exchange (SCE) level and shows hypersensitivity towards monofunctional alkylating agents and bromodeoxyuridine (BrdUrd). The 2 cell clones have been transfected with and allowed to incorporate stable genomic inserts of the vector pIII-14gpt, which contains 2 tandemly arranged neo gene fragments with a common 400-bp region of homology. Recombination between the truncated neo genes gives rise to geneticin sulfate (G 418)-resistant revertants with a spontaneous frequency of about 10(-4) in both cell clones. In CHO:5 an increased frequency of revertants was obtained after treatment with methyl methanesulfonate (MMS) and mitomycin C (MMC), while HN2, benz[a]pyrene diolepoxide (BPDE) and X-irradiation gave negative results. EM9:2 showed about the same increase of revertants after treatment with MMS as CHO:5, but in a 10-fold lower dose range. HN2 as well as BrdUrd induced revertants in EM9:2. These results show that mutagenic agents (MMS, MMC, HN2, BrdUrd) can induce homologous recombination in this system. This effect does not seem to be an unspecific effect of DNA damage (no effect of X-ray and BPDE), or related to SCE induction in general (similar spontaneous and MMS-induced frequencies of revertants in CHO:5 and EM9:2). However, the positive effect of BrdUrd in EM9:2 and the difference between CHO:5 (negative) and EM9:2 (positive) with regard to HN2-induced revertants suggest that certain types of DNA damage are more recombinogenic in EM9 than in 'normal' CHO cells, which possibly reflects the specific mutation in the former cell line.
Hellgren
D
D
Department of Clinical Genetics, Karolinska Institute, Stockholm, Sweden.
Sahlén
S
S
Lambert
B
B
D000818:Animals; D002460:Cell Line; D004260:DNA Repair; D004351:Drug Resistance / Q000235:genetics; D009153:Mutagens / Q000494:pharmacology*; D009154:Mutation; D009355:Neomycin / Q000494:pharmacology*; D010957:Plasmids; D011995:Recombination, Genetic / Q000187:drug effects* / Q000528:radiation effects; D012854:Sister Chromatid Exchange; D014162:Transfection; D014965:X-Rays
D009153:Mutagens; D009355:Neomycin