A Drosophila RNA Polymerase II Transcription Factor Contains a Promoter-region-specific DNA-binding Activity

Drosophila RNA polymerase II requires at least two chromatographically distinct transcription factors (designated A and B) to initiate transcription accurately in vitro. We describe the partial purification and concentration of one of these transcription factors, the B factor. Footprint analysis of the B fraction demonstrated the presence of a sequence-specific DNA-binding component in the transcription factor preparation. This component binds specifically to a 65 bp region of DNA surrounding the start point of transcription of the histone H3, H4, and actin 5C genes. Included in this binding region is the TATA box, the start point of transcription, and a portion of the leader region. The pattern of protection from DNAase I cleavage on the coding strand of the histone H3 gene is asymmetric with regard to the complementary noncoding strand. Sequence-specific binding of the B fraction occurs in the apparent absence of RNA polymerase II. The potential function of the binding component in the initiation of transcription by RNA polymerase II is discussed.