Promotion of specific in vitro transcription by excised "TATA" box sequences inserted in a foreign nucleotide environment

Abstract

We have cloned into plasmid pBR322 a DNA fragment extending from position -32 to position -12 of the adenovirus type 2 major late promoter region (position +1 referring to the cap site). In vitro transcription experiments show that this 21 base pair sequence, which contains the Goldberg-Hogness or "TATA" box, is both necessary and sufficient for specific initiation of transcription by RNA polymerase B (or II). Furthermore, we show that similar sequences, randomly occurring in the bacterial plasmid pBR322, are also recognized by the RNA polymerase B transcription machinery and able to promote specific in vitro transcription. Finally, we discuss the possible importance of the nucleotide sequence of the start region in the actual efficiency of initiation of in vitro transcription.