Adeno-Associated Virus Rep78 Binds to E2-Responsive Element 1 of Bovine Papillomavirus Type 1

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Abstract
Adeno‐associated virus type‐2 (AAV‐2) is a helper‐dependent parvovirus that has been implicated in the inhibition of replication and oncogenic transformation of bovine papillomavirus type‐1 (BPV‐1) and other transforming DNA viruses. Previous studies have suggested that the Rep78 protein of AA V‐2 is a key player mediating this effect. In this report we have analyzed the effect of AAV‐2 Rep78 protein on the regulation of gene expression of a reporter gene under the control of the long control region (LCR) of BPV‐1. Our results show that Rep78 is capable of down‐regulating the promoter activity of the LCR in vivo in tissue culture cells. Inhibition of LCR activity in vivo suggested the need for Rep78 to bind to a region of the LCR promoter spanning the E2‐responsive elements of BPV‐1. This observation was further confirmed in vitro with gel shift assays showing specific binding of Rep78 to DNA oligonucleotides containing E2‐responsive element 1 (E2RE1) sequences of BPV‐1 LCR. Our results expand the understanding of the mechanism of trans‐regulation mediated by Rep78 and involving this protein and DNA sequences with complex secondary structure.

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