Induced accumulation of polyubiquitin gene transcripts in HeLa cells after UV-irradiation and TPA-treatment

There are three species of ubiquitin gene transcripts in HeLa cells, termed UbA (approximately 0.7 kb), UbB (approximately 1.1 kb) and UbC (approximately 2.5 kb). In the present report, the UbC transcript was shown to accumulate up to 2.5-fold after irradiation with UV light or treatment with the phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA). The kinetic analysis indicated that the induced accumulation of UbC was rapid and transient; maximal accumulation of UbC was induced at 2.5 h after UV irradiation or 3 h after TPA treatment. Inhibition of a de novo protein synthesis by cycloheximide did not repress the induction of UbC after treatment with UV light and TPA. On the other hand, induction of UbA and UbB, in most cases, was not observed. UV-inducibility of human ubiquitin conjugating enzyme, E2(17k), was also tested. E2(17k) is a protein with high sequence similarity to the product of yeast DNA repair gene, RAD6. While the RAD6 gene has been reported to be inducible by UV light, no change in E2(17k) gene transcript was observed after UV irradiation.