p53 is involved in regulation of the DNA repair gene O⁶-methylguanine-DNA methyltransferase (MGMT) by DNA damaging agents

Abstract

The DNA repair protein O⁶-methylguanine-DNA methyltransferase (MGMT) is inducible by genotoxic stress. MGMT induction results from transcriptional activation of the MGMT gene which is a specific response to DNA damage. A possible factor involved in triggering MGMT induction might be p53, because both p53 and MGMT are activated by DNA breaks. To study the effect of p53 on induction of the MGMT gene, we compared the presence of functional wild-type (wt) and mutant p53 with MGMT expression level in various mouse fibroblasts and rat hepatoma cell lines upon genotoxic treatment. Cells which responded to ionizing radiation (IR) by MGMT induction displayed functional p53, whereas in cells not expressing wt p53, MGMT induction was not observed. Also, the cloned MGMT promoter was inducible by IR upon transfection into p53 wt cells, but not in cells deficient for p53. Thus, expression of wt p53 appears to be required for induction of MGMT mRNA and protein by IR. On the other hand, transfection of a MGMT-promoter-CAT construct together with p53 (either wt or mutant) in cells expressing wt p53 markedly reduced the basal activity of the MGMT promoter whereas cotransfection with a p53 antisense construct slightly increased MGMT promoter activity. Furthermore, cotransfection of MGMT promoter with wt or mutant p53 in p53 wt cells reduced radiation evoked MGMT promoter induction. Thus, transfection mediated high level expression of p53 has inhibitory effect both on basal MGMT promoter activity and its activation by IR. The results give evidence for involvement of p53 in DNA damage-induced MGMT promoter activation.