Identification of baculoviral factors required for the activation of enhancer-like polyhedrin upstream (pu) sequence.

Previously, we identified a novel enhancer-like element, the polyhedrin upstream (pu) sequence, in the genome of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV), which activates several early promoters. The activation requires co-infection of AcMNPV, suggesting that viral gene products are needed for pu-mediated promoter activation. DNA replication assay showed that the pu sequence did not assist in DNA replication and suggested its involvement in activated transcription from target promoters. In order to identify the viral genes responsible for pu-dependent activation of early promoters, a set of overlapping cosmid clones covering the entire 134-kb AcMNPV genome were constructed and screened. Our results identified three viral genes ie1, ie2, and pe38, which function in concert with pu to activate target promoters. In addition, these three viral factors can substitute for the entire virus for the synergistic promoter activation mediated by pu and the known baculovirus enhancer, the homologous region.