Transcription of adenovirus 2 major late and peptide IX genes under conditions of in vitro nucleosome assembly.

Abstract

Plasmid DNA containing adenovirus 2 major late and peptide IX genes was assembled into nucleosomes in vitro, and the assembled nucleosomes were used as a template to study the regulatory mechanism of transcription initiation under these conditions. Neither the major late nor peptide IX genes was transcribed on the already-assembled nucleosomes. However, the major late gene, but not the peptide IX gene, was transcribed efficiently when the DNA was incubated with HeLa cell extracts prior to assembly into nucleosomes. These results indicate that prebinding of some component in the cell extracts to DNA is essential to activate transcription of the major late gene on nucleosomes assembled under the conditions used here. Since gene IX on the nucleosomes was not transcriptionally active regardless of preincubation of DNA with the extracts, some other component or another, different template structure which is not able to be identified in an in vitro system with deproteinized DNA template might be required for activation of peptide IX gene transcription. To know the function of the upstream sequences of the major late gene, effects of the deletion on transcription of nucleosomes were compared with that of deproteinized DNA. The result showed that depression of transcription by deleting the upstream sequences had more effect on nucleosomes than on deproteinized DNA.