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An Element Binding a C/EBP-related Transcription Factor Contributes to Negative Regulation of the Bovine Papillomavirus Type 4 Long Control Region

The Beatson Institute for Cancer Research, CRC Beatson Laboratories, Garscube Estate, Switchback Road, Bearsden, Glasgow G61 1BD, U.K.

Deletion of the NR2 element of the long control region (LCR) of bovine papillomavirus type 4 (BPV-4) was observed previously to lead to a fivefold increase in enhancer activity of a subfragment of the LCR. Further characterization of this element indicates that mutations in NR2 lead to increased enhancer activity in both mouse CT3 fibroblasts and in a transformed bovine epithelial cell line derived from an alimentary canal papilloma/in situ carcinoma, but not in primary bovine keratinocytes. Since similar oligonucleotide-nuclear factor complexes were obtained in electrophoretic mobility shift assays (EMSA) for all three cell types, the observed difference in negative activity may result from variation in the NR2-binding factor itself between primary and established/transformed cell lines, or from the involvement of other factors that vary between the lines. Characterization of the NR2-binding factor by heat stability and antibody supershifts in EMSA indicate that the factor is related to the CCAAT/enhancerbinding protein (C/EBP) family, and that one component of the complexes may be C/EBP.

Received 26 April 1994; accepted 22 June 1994.

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