Abstract
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.
Highlights
The papillomavirus E2 open reading frame encodes several proteins that bind to the consensus E2-binding site (E2BS), ACCN6GGT, and regulate viral transcription and DNA replication [8, 9]
FLAG-protein-associated factor (p/CAF) did not co-precipitate with glutathione S-transferase (GST) along with FLAG-p/CAF or GST-E2 alone, indicating that p/CAF binds to human papillomavirus (HPV)-18 E2 in vivo
These results showed that p/CAF binds to HPV-18 E2 in mammalian cells
Summary
The papillomavirus E2 open reading frame encodes several proteins that bind to the consensus E2-binding site (E2BS), ACCN6GGT, and regulate viral transcription and DNA replication [8, 9]. Its ability to stimulate transcription is independent of p300/CBP, but is dependent on the intrinsic HAT activity of p/CAF [3]. Incubation of the transfected cell extracts with GST or protein G (detecting for FLAG antibody) resin resulted in the co-precipitation of GST-18E2 and FLAG-p/CAF. To determine whether p/CAF binds to HPV-18 E2 in vivo, we co-transfected GST-fused HPV-18 E2 and FLAG-tagged p/CAF expression vectors into 293T cells.
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