title: Characterization of the DNA-binding activity of the E1 and E2 proteins and the E1/E2 complex of human papillomavirus type 33.
abstract: The E1 and E2 proteins of papillomaviruses are essential for the initiation of viral DNA replication. We have purified the E2 protein of human papillomavirus type 33 (HPV-33) by immunoaffinity chromatography. The purified E2 protein bound with high affinity to all four consensus binding sites of HPV-33 (Kd approximately equal to 2 x 10(-10)M). A putative E2 binding site differing at one position in the second stem of the palindrome was not bound by E2. The E1 protein of HPV-33 purified by affinity chromatography using glutathione S-transferase as tag displayed specific DNA-binding activity in footprint analyses protecting HPV-33 nucleotides 7896 to 7909/1 to 18 from DNasel digestion. Hypersensitive sites at position 6 on the sense and position 1 on the antisense strand were observed in the middle of the protected region. An E1/E2 complex protected the E1 binding site and E2 binding sites from DNasel digestion suggesting that both proteins retain DNA-binding activity in the complex.
authors.affiliation: Institut für, Medizinische Mikrobiologie und Hygiene, Hochhaus am Augustusplatz, Universität Mainz, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Müller
authors.forename: T
authors.initials: T
authors.lastname: Giroglou
authors.forename: M
authors.initials: M
authors.lastname: Sapp
mesh_terms: D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000378:metabolism*; D027383:Papillomaviridae / Q000378:metabolism*; D011485:Protein Binding; D014759:Viral Envelope Proteins / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D009856:Oncogene Proteins, Viral; D014759:Viral Envelope Proteins