title: Characterization of the non-specific DNA binding properties of the Adenoviral IVa2 protein.
abstract: Human Adenovirus (Ad) is a non-enveloped, icosahedral virus with a linear, double-stranded DNA genome. The Ad IVa2 protein is involved in multiple viral processes including viral late gene transcription and virus assembly. Previous studies have shown that IVa2 loads additional viral proteins onto conserved DNA elements within the Ad genome to regulate these viral processes. IVa2 also possesses strong non-specific DNA binding activity, and it is likely it uses this activity to recruit proteins to the conserved DNA elements. Here we have investigated the non-specific DNA binding activity of IVa2 using nitrocellulose/DEAE filter binding and sedimentation equilibrium techniques. We have analyzed our data using the McGhee and Von Hippel approach [1], and find that IVa2 binds with strong, positive nearest-neighbor cooperativity. In addition, we describe how to apply the McGhee and von Hippel approach to directly analyze sedimentation equilibrium data using non-linear least-squares methods. We discuss the implications of these results with respect to current virus assembly mechanisms.
authors.affiliation: University of Colorado Denver, School of Pharmacy, Dept. Pharm. Sciences, C238 12850 E. Montview Blvd., V20-4121, Aurora, CO 80045.
authors.forename: Teng-Chieh
authors.initials: TC
authors.lastname: Yang
authors.affiliation: University of Colorado Denver, School of Pharmacy, Dept. Pharm. Sciences, C238 12850 E. Montview Blvd., V20-4121, Aurora, CO 80045; Alliance Protein Laboratories, 6042 Cornerstone Ct West A, San Diego, CA 92121. Electronic address: Karl.Maluf@ap-lab.com.
authors.forename: Nasib Karl
authors.initials: NK
authors.lastname: Maluf
mesh_terms: D000260:Adenoviruses, Human / Q000737:chemistry* / Q000378:metabolism; D004279:DNA, Viral / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013816:Thermodynamics; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics
chemical_list: D004279:DNA, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; C056232:iva2 protein, adenovirus
keywords: Analytical ultracentrifugation; IVa2; McGhee and von Hippel non-specific binding model; Non-specific DNA binding; Sedimentation equilibrium; Virus assembly