A 12,500 MW protein is coded by region E3 of adenovirus

There is an open reading frame between ATG291 and TGA612 in the early region E3 transcription unit of adenovirus 2 (Ad2) that could encode a protein of 12,500 MW (12.5K). To address whether this protein is synthesized, we generated an antiserum against a TrpE-12.5K fusion protein which was expressed in Escherichia coli. This antiserum immunoprecipitated a doublet of about 12.5K apparent MW from [35S]Cys-labeled cells infected with Ad2, Ad5, and various mutants in other E3 genes. Mutants in the 12.5K gene did not produce this protein, and an in-frame deletion mutant showed a protein with a corresponding decrease in size. Cell-free translation of hybridization-purified RNA indicated that 12.5K is coded by E3 mRNA i. mRNA i is relatively scarce, and 12.5K is synthesized in correspondingly small amounts. The 12.5K protein was synthesized at early and late stages of infection in comparable amounts. Pulse-chase experiments indicated that 12.5K has a half-life of about 10 hr. The function of 12.5K is unknown, and the 12.5K gene can be deleted without affecting virus growth in cell culture. However, 12.5K is likely to be important in vivo because the gene is highly conserved in both Ad2 and Ad5 (group C adenoviruses), and also in Ad3 (group B).