The bovine papillomavirus P2443 promoter is E2 trans-responsive: evidence for E2 autoregulation.

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RESUMO

The bovine papillomavirus type 1 (BPV-1) P2443 promoter is located just upstream of the E2, E3, E4 and E5 open reading frames (ORFs) and is active in both transformed rodent cells and in productively infected warts. Analysis of viral RNA structures suggests that transcripts from this promoter encode the E2 transactivator as well as the E5 oncoprotein. To study expression of P2443, the chloramphenicol acetyltransferase (CAT) reporter gene was placed downstream of this promoter, deleting the E2 and E5 ORFs, in a plasmid which contained all BPV-1 upstream sequences including the long control region (LCR). By itself, this plasmid demonstrated a low level of activity in transient assays and could be transactivated to a high level by the full-length E2 product. Transactivation of P2443 expression by E2 required the LCR in cis in an orientation- and position-independent manner, suggesting that this transactivation was mediated through the E2 responsive enhancer elements (E2RE) located within the LCR. Primer extension analysis of the 5' ends of the viral transcripts from pooled cells expressing these P2443/CAT plasmids confirmed that E2 transactivation results in an increase in the steady-state levels of RNA initiated from the P2443 promoter. Furthermore, E2 transactivation of the P2443 promoter could be inhibited by the trans-repressor encoded by the 3' portion of the E2 ORF. Thus, expression of the E2 transactivator and the E5 oncoprotein is directly regulated by transcriptional factors encoded by the E2 ORF.

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