In human and mouse cell lines, expression of exogenous genes was enhanced by treatment with 2-aminopurine (2-AP). Chloramphenicol acetyltransferase (CAT) and neomycin phosphotransferase activities were increased by up to 50-fold upon 2-AP treatment of cells permanently transfected with genes encoding these enzymes. Neomycin phosphotransferase activity was also increased by this treatment in cells infected with a retroviral vector carrying the neomycin phosphotransferase gene. 2-AP-mediated increase in CAT activity was observed in various cell lines which had been permanently transfected with chimeric CAT genes containing transcriptional regulatory elements of the interferon-inducible genes 6-16 and 561, SV40 early genes, mouse mammary tumor viral gene, or metallothionein II gene. The increase in the cellular CAT enzymatic activity was due to an elevated level of CAT protein. The 2-AP-mediated enhancement of CAT expression was operative at the translational level; the rate of transcription of CAT mRNA or its steady-state level was affected only marginally. The translational up-regulation by 2-AP was restricted to the genes introduced from outside; there was no gross change in the rate of synthesis of other cellular proteins.