Adenoviral early region 4 is required for efficient viral DNA replication and for late gene expression

Abstract

H2dl808 is a defective deletion mutant of human adenovirus 2 lacking most of transcriptional early region 4. Although the mutant can be grown in the complementing cell line W162, it is defective in human cell lines normally used to propagate adenovirus. In such nonpermissive cells, H2dl808 exhibits a severe defect in late gene expression, accumulating very small amounts of viral late messages and producing correspondingly small amounts of viral late proteins. H2dl808 also exhibits a defect in viral DNA synthesis: 24 h after infection, H2dl808-infected nonpermissive cells contain five- to sevenfold less viral DNA than those cells infected with wild-type adenovirus. H2dl808-infected nonpermissive cells eventually accumulate a significant amount of viral DNA. However, the rate of synthesis of viral proteins late in mutant infection remains much lower than that observed in wild-type infection at a time when DNA accumulation is comparable. Thus, the mutant's late protein synthesis defect is probably not due solely to its reduced accumulation of viral DNA. Finally, H2dl808 is much less efficient than wild-type virus in the inhibition of host cell protein synthesis in infections of nonpermissive cells. These observations imply roles for early region 4 products in several aspects of the viral growth cycle, including DNA replication, late gene expression, and host cell shutoff.