Differential activation of the E2F transcription factor by the adenovirus EIa and EIV products in F9 cells

Expression studies of the early EIIa transcription unit (EIIaE) of the adenovirus EIa-deletion mutant dl312 in murine embryonal carcinoma stem cells suggested that these cells contain an activity that substitutes for the viral EIa. To further characterize this cellular EIa-like activity, we analyzed expression of the EIIaE promoter as well as the binding activity of the cognate E2F transcription factor after infection of F9 embryonal carcinoma cells and their differentiated derivatives with wild-type adenovirus, EIa (dl312), or EIV (dl808) deletion mutants. We show that, in contrast to the viral EIa proteins that transactivate the EIIaE promoter in F9 cells only after differentiation, the viral EIV products activate the EIIaE promoter most efficiently in undifferentiated F9 cells. We also show that the EIV products induce a specific modification of the E2F transcription factor leading to its cooperative binding to the EIIaE promoter. Although the EIa-dependent transactivation of EIIaE in differentiated cells is also in part mediated by E2F, it does not by itself correlate with the simultaneous binding of two E2F molecules. In these cells E2F dimer binding only occurs as a secondary effect of EIa that also stimulates EIV expression. Our results suggest that EIa and EIV act through separate pathways, inversely regulated during cell differentiation, with the so-called "EIa-like" activity contributing to this modulation.