The specific DNA recognition sequence of the bovine papillomavirus E2 protein is an E2-dependent enhancer.

P. Hawley-Nelson, Elliot Androphy, D. R. Lowy, J. T. Schiller

Research output: Contribution to journalArticle

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Abstract

The upstream regulatory region (URR) of the bovine papillomavirus (BPV) genome contains an enhancer that is activated by a BPV E2 gene product. We have previously found that a bacterially derived E2 fusion protein specifically interacted with several fragments of URR DNA, suggesting that E2 may activate transcription by directly binding to the enhancer. Each of the bound fragments contains at least one copy of a conserved motif (ACCN6GGT). To determine if this motif is required and sufficient for specific E2 binding, we have now constructed a bacterial expression vector that encodes a full-length E2 peptide and developed a refinement of the McKay DNA immunoprecipitation assay that allows the determination, to the nucleotide level, of the minimum sequence required for specific binding. The results show that the E2 recognition sequence is a single copy of this motif and that the variant ACCGN4CGGT is bound with greater affinity than the minimum ACCN6GGT motif. An oligonucleotide encoding the motif was able to inhibit E2-dependent trans-activation in a transient transfection assay, indicating that the virally encoded E2 also interacts with this sequence in mammalian cells. When present in two or more copies, but not in a single copy, the E2 binding element had intrinsic enhancer activity but only in cells expressing E2. The results indicate that the conserved motif alone is sufficient for E2-mediated enhancement and that the binding of E2 to the motif is probably required for efficient enhancement. Since a single motif did not have a significant enhancer activity, it is likely that bound E2 molecules act cooperatively in activating transcription.

Original languageEnglish (US)
Pages (from-to)525-531
Number of pages7
JournalEMBO Journal
Volume7
Issue number2
StatePublished - Feb 1988
Externally publishedYes

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Nucleic Acid Regulatory Sequences
Transcription
Assays
Genes
Cells
DNA
Immunoprecipitation
Oligonucleotides
Transfection
Fusion reactions
Nucleotides
Chemical activation
Genome
Peptides
Molecules
Proteins
Bovine papillomavirus E2 protein

ASJC Scopus subject areas

  • Cell Biology
  • Genetics

Cite this

The specific DNA recognition sequence of the bovine papillomavirus E2 protein is an E2-dependent enhancer. / Hawley-Nelson, P.; Androphy, Elliot; Lowy, D. R.; Schiller, J. T.

In: EMBO Journal, Vol. 7, No. 2, 02.1988, p. 525-531.

Research output: Contribution to journalArticle

Hawley-Nelson, P. ; Androphy, Elliot ; Lowy, D. R. ; Schiller, J. T. / The specific DNA recognition sequence of the bovine papillomavirus E2 protein is an E2-dependent enhancer. In: EMBO Journal. 1988 ; Vol. 7, No. 2. pp. 525-531.
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