Irradiation-induced expression of O6-methylguanine-DNA methyltransferase in mammalian cells.
O6-Methylguanine-DNA methyltransferase (MGMT) is a DNA repair protein which plays an important role in chemotherapy, mutagenesis, and carcinogenesis. The specific activity of MGMT in female rat liver can be induced by approximately 20-fold by treatment of the rats with gamma-irradiation. Maximum response occurred 48 h after 15 Gy irradiation. MGMT levels in male rats were induced by only 3-fold. MGMT activity was also induced by irradiation of rat hepatoma H4IIE cells with a 3-fold increase noted after treatment with 3 Gy. Northern analysis and nuclear run-on assays indicated that the induction of MGMT was regulated at the transcriptional level. The radiation-mediated increase in MGMT was blocked by H7, a protein kinase inhibitor, but not by H89, an inhibitor of protein kinase A. Hydroxyl radicals may play a role in the induction mechanism since dimethyl sulfoxide, a radical scavenger, blocked the radiation-mediated increase in MGMT. MGMT activity was also increased by treatment of the cells with H2O2, in accordance with the involvement of activated oxygen species in the induction of MGMT. Finally, the addition of cycloheximide, an inhibitor of protein synthesis, prior to but not after irradiation, abolished the increase in MGMT activity.
Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, New Hampshire 03756.
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Chan
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Wu
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Eastman
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Bresnick
D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000502:physiology / Q000528:radiation effects; D003513:Cycloheximide / Q000494:pharmacology; D015342:DNA Probes; D004307:Dose-Response Relationship, Radiation; D004790:Enzyme Induction / Q000528:radiation effects; D005260:Female; D005720:Gamma Rays; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology / Q000528:radiation effects*; D008114:Liver Neoplasms, Experimental; D008297:Male; D008780:Methyltransferases / Q000096:biosynthesis* / Q000235:genetics / Q000528:radiation effects; D019853:O(6)-Methylguanine-DNA Methyltransferase; D009928:Organ Specificity; D011061:Poly A / Q000235:genetics / Q000302:isolation & purification / Q000528:radiation effects; D012313:RNA / Q000235:genetics / Q000302:isolation & purification / Q000528:radiation effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012727:Sex Characteristics
D015342:DNA Probes; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D003513:Cycloheximide; D008780:Methyltransferases; D019853:O(6)-Methylguanine-DNA Methyltransferase