Here we analyzed the effect of the suppressor proto-oncogene p53 on transcription from the P2 promoter of the murine c-myc gene. c-myc promoter constructs were coupled to the chloramphenicol acetyl-transferase (CAT) gene and were transiently transfected into a human glial cell along with plasmids overexpressing wild-type or mutant p53. It was found that significant repression of c-myc transcription took place following cotransfection with wild-type but not mutant p53. However wild-type p53 did not suppress transcription from the SV40 early promoter or from the MHC promoter. Promoter-CAT constructs containing only the ME1a2 or E2F elements, from the P2 promoter, were repressed by p53, indicating that p53 may exert its effect at these two sites within the P2 promoter. Finally, when the SV40 T antigen and wild-type p53 were expressed together in glial cells the repressive effect of p53 was abolished.