Effect of transfection of human poly(ADP-ribose)polymerase in Chinese hamster cells on mutagen resistance.
Poly(ADP-ribose)polymerase (PARP) is a DNA-binding protein that is activated upon induction of DNA breaks and supposed to play a role in DNA repair. To elucidate the effect of overexpression of PARP on the resistance of cells to mutagens, Chinese hamster ovary cells (both the line CHO-9 and the mutagen-hypersensitive derivative 27-1) were transfected with the human PARP cDNA along with pSV2neo. Treatment of the transfected cell population with a high dose of MNNG and selection with G418 gave rise to a significant increase of neo+ clones, as compared to the control transfection with pSV2neo + salmon sperm DNA. The frequency of survivors in these mass culture experiments was lower, however, than after transfection with the bacterial ada gene encoding the DNA repair protein O6-alkylguanine-DNA alkyltransferase. Thus transfection of PARP cDNA in CHO cells is only weakly effective in inducing alkylation resistance. This was confirmed by analyzing the mutagen resistance of individual PARP transfectant clones derived from CHO-9 and 27-1 cells that expressed increased levels of PARP mRNA, protein and PARP activity. These strains were slightly more resistant to the toxic effect of MMS and showed a reduced frequency of MMS-induced chromosomal aberrations. CHO-9-PARP transfectants also gained resistance to UV. From these data we conclude that, in CHO cells, PARP is limiting in handling critical lesions during the repair process and that increase of the amount of PARP protein can elicit some protection against genotoxic effects of mutagens.
Fritz
G
G
Nuclear Research Center Karlsruhe, Department of Genetics, Germany.
Auer
B
B
Kaina
B
B
D000818:Animals; D016587:Antimutagenic Agents; D015153:Blotting, Western; D016466:CHO Cells; D002470:Cell Survival; D002869:Chromosome Aberrations; D006224:Cricetinae; D004260:DNA Repair; D006801:Humans; D008741:Methyl Methanesulfonate / Q000633:toxicity; D008769:Methylnitronitrosoguanidine / Q000633:toxicity; D008780:Methyltransferases / Q000378:metabolism; D019853:O(6)-Methylguanine-DNA Methyltransferase; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D014162:Transfection
D016587:Antimutagenic Agents; D008769:Methylnitronitrosoguanidine; D008741:Methyl Methanesulfonate; D008780:Methyltransferases; D019853:O(6)-Methylguanine-DNA Methyltransferase; D011065:Poly(ADP-ribose) Polymerases