Expression of human O-Methyl Guanine Methyl Transferase (MGMT) in Post Replication Repair (PRR) deficient CHO-UV-1 cells: Compensation for hypersensitivity to methylating and ethylating agents but not to mitomycin C

The cDNA for human MGMT was transfected into and expressed in CHO cells and the post-replication repair deficient mutant CHO-UV-1 cell, both of which are devoid of endogenous MGMT activity. Expression of MGMT activity was demonstrated by measurement of activity and by imiminoblot analysis. The mutant phenotype of UV-1 is characterized by extreme hypersensitivity to killing by methylating and ethylating agents as well as the antitumor antibiotic mitomycin C (MMC). MGMT expression conferred equivalent, supra-normal levels of resistance to killing by MNNG (N-methyl-N′-nitro-nitrosoguanidine) or EMS (ethyl methanesulfonate) on CHO and UV-1, but had no effect on the lethality of MMC. So, even though a mutated gene other than MGMT is known to underlie the pleiotropic phenotype of UV-1, expression of MGMT compensates for part of this phenotype. This result indicates that attempts to concordance map and clone the gene(s) responsible for the UV-1 phenotype can be complicated when using MNNG selection due to compensation by the MGMT gene. These results also indicate that the post-replication repair deficient phenotype characterized in CHO-UV-1 cells, will be masked in cells normally expressing MGMT due to MGMT-mediated resistance to methylating and ethylating agents. © 1997 Plenum Publishing Corporation.