The acidic transcriptional activation domains of VP16 and p53 bind the cellular replication protein A and stimulate in vitro BPV-1 DNA replication.
For papillomavirus DNA replication, the E2 enhancer protein cooperatively assists in binding of the E1 helicase to the origin. We report that, at limiting E1 and E2 levels, the enhancer proteins GAL4-VP16 and GAL4-p53(1-73) stimulate BPV in vitro DNA replication. This cell-free system was used to ascertain whether the acidic activation domains have a cellular target important for replication. Cellular extracts were depleted of replication activity by passage through a VP16 affinity column. The protein depleted was the cellular factor replication protein A. The direct interaction between replication protein A and VP16, as well as the activation of replication by VP16, is dependent upon the C-terminus of the VP16 activation domain. E2 and the activation domain of p53 also interact with replication protein A. We suggest that a link between transcription and replication involves factors that help convert a closed DNA complex to an open complex.
Department of Molecular and Cell Biology University of California, Berkeley 94720.
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D017502:Adenovirus E2 Proteins; D052688:Bovine papillomavirus 1 / Q000378:metabolism*; D002474:Cell-Free System; D004261:DNA Replication; D004279:DNA, Viral; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D011485:Protein Binding; D051716:Replication Protein A; D015534:Trans-Activators / Q000378:metabolism*; D014158:Transcription, Genetic
D017502:Adenovirus E2 Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C070633:Gal-VP16; D051716:Replication Protein A; D015534:Trans-Activators