E1A gene products stimulate in vitro transcription from the adenovirus early region 4 promoter by enhancing a stable preinitiation complex

Abstract

A cell-free transcription system using nuclear extracts prepared from adenovirus type 5 (Ad5)-infected and mock-infected cells were utilized to study the E1A-mediated transactivation of transcription from the Ad5 early region 4 (E4) promoter. The transcription activity of Ad-infected cell extracts was several times higher than that of mock-infected cell extracts when the E4 wild-type (WT) promoter was used. The increased activity required expression of E1A proteins. However, the transcription activities of both cell extracts were not different when the mutant promoter containing only the E4 TATA box was used. The formation of a stable preinitiation complex at the E4 WT promoter was facilitated by expression of E1A, whereas it was not facilitated at the mutant promoter. The results suggested that the E1A proteins stimulated transcription from the E4 promoter by enhancing the formation of a stable preinitiation complex through the upstream elements, together with the TATA box.