A 14,500 MW protein is coded by region E3 of group C human adenoviruses.

There is an ORF in the early region E3 transcription unit of human adenovirus 5 (Ad5) which could encode a protein of 14,500 MW (14.5K). This ORF is conserved in Ad5 and Ad2, both group C adenoviruses, and also in Ad3 and Ad7, both group B adenoviruses. To address whether the 14.5K protein is synthesized, we prepared antisera against synthetic peptides corresponding to residues 19-34 or 118-132 in the Ad5 version of 14.5K, and also against a TrpE-14.5K fusion protein expressed in Escherichia coli. These antisera immunoprecipitated the [35S]Met-labeled 14.5K protein from KB cells infected with rec700 (an Ad5-Ad2-Ad5 recombinant), Ad2, and a variety of E3 mutants. Mutants in the 14.5K ORF did not produce the 14.5K protein. The 14.5K is coded in large part, although probably not exclusively, by E3 mRNA f, as indicated by immunoprecipitation of 14.5K from cells infected with mutants that overproduce or underproduce mRNA f. The 14.5K migrated as five to six bands on SDS-PAGE after immunoprecipitation or Western blot, suggesting that it undergoes post-translational modification. Two bands of 14.5K were obtained by cell-free translation of 14.5K from mRNA purified by hybridization from infected cells.