title: Cooperative binding of the E2 protein of bovine papillomavirus to adjacent E2-responsive sequences.
abstract: The DNA-binding properties of purified full-length E2 protein from bovine papillomavirus type 1 have been investigated by utilizing a quantitative gel shift analysis. By using a recombinant baculovirus which express the E2 open reading frame from the polyhedrin promoter, the full-length E2 protein was synthesized in insect cells and purified to homogeneity by using an E2 binding site (ACCGN4CGGT)-specific oligonucleotide column. The Kd of E2 binding to a 41-bp oligonucleotide containing a single binding site was found to be 2 x 10(-11) M. When two binding sites were included on an oligonucleotide, cooperative binding to these sites by the E2 protein was observed. A cooperativity parameter of 8.5 was determined for E2 binding to two sites. An 86-amino-acid peptide encompassing the C terminus of the protein retains the ability to bind E2 binding sites with a Kd of 4 x 10(-10) M but exhibits slight cooperativity of binding to two adjacent sites. A major determinant for cooperative binding of the full-length E2 protein is thus encoded by the N-terminal amino acids outside the minimal DNA binding domain.
authors.affiliation: Howard Hughes Medical Institute, University of Chicago, Illinois 60637.
authors.forename: P
authors.initials: P
authors.lastname: Monini
authors.forename: S R
authors.initials: SR
authors.lastname: Grossman
authors.forename: B
authors.initials: B
authors.lastname: Pepinsky
authors.forename: E J
authors.initials: EJ
authors.lastname: Androphy
authors.forename: L A
authors.initials: LA
authors.lastname: Laimins
mesh_terms: D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D052688:Bovine papillomavirus 1 / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D015870:Gene Expression; D008969:Molecular Sequence Data; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; C051550:E2 protein, Bovine papillomavirus; D014764:Viral Proteins