Transcriptional factors have been implicated in eukaryotic DNA replication. We have studied the potential function of a viral promoter sequence in DNA replication. The hepatitis B virus (HBV) pregenomic promoter is regulated by two enhancers and cis-elements. The G-C rich region between 1734-1754 nt, which contains two SP1 binding sites, is necessary for transcription origin and HBV replication. We found that the Abf1-binding B3 element in yeast ARS1 can be functionally replaced by the viral Sp1-binding DNA sequence, which activates transcription from the HBV C promoter. Further, yeast RAP1 bound to the viral Sp1 binding sites in vitro. These results suggest that RAP1 binds to the Sp1 binding sites and stimulates yeast DNA replication.