E1A expression might be controlled by miR-214 in cells with low adenovirus productivity.

In this study, we explored the differences in the human adenovirus type 5 (Ad5) production efficiencies of various cell types. The rate of virus production was higher in several cell lines, such as HeLa cells, than in Saos-2 cells. The expression level of the coxsackie and adenovirus receptor (CAR) protein, an adenovirus receptor, was very similar among these cell lines. Although no significant difference in the expression of early region 1A (E1A) mRNA was detected, the amount of E1A protein in the Saos-2 cells was markedly lower than that in HeLa cells. Proteasome inhibitor treatment did not rescue the quantity of E1A in the Saos-2 cells, suggesting that their decreased E1A protein expression is not due to protein decay. To examine the different expression of E1A protein, we employed a bioinformatics approach to identify miRNA that target the 3'-untranslated region (3'-UTR) of E1A mRNA and identified miR-214 as a highly promising candidate. In Saos-2 cells, which have abundant levels of endogenous miR-214, the expression of luciferase was dramatically repressed, when the reporter gene was fused with the 3'-UTR of E1A mRNA including an miR-214 binding site. On the other hand, the activity from the same reporter was unchanged in HeLa cells, which display low-level miR-214 expression. Finally, we confirmed that the knockdown of the miR-214 upregulated the productive efficiency of the virus. These findings indicate that cellular miR-214 is capable of inhibiting adenovirus replication by regulating the translation of E1A protein.