p300/CREB-binding protein-associated factor (p/CAF), a transcriptional co-activator, interacts with co-activator p300/CBP and acidic transcription factors. p/CAF mediates transcriptional activation by acetylating nucleosomal histones and cellular factors. Previously we reported that CBP binds to human papillomavirus E2 and activates E2-dependent transcription (Lee, D., Lee, B., Kim, J., Kim, D. W., and Choe, J. (2000) J. Biol. Chem. 275, 7045-7051). Here we show that p/CAF binds to the human papillomavirus E2 protein in vivo and in vitro and activates E2-dependent transcription. CBP along with p/CAF synergistically activates E2-dependent transcription. In addition, the histone acetylase activity of p/CAF is required for efficient activation of E2 transcriptional activity. These results suggest that p/CAF is a transcriptional co-activator of the human papillomavirus E2 protein.