Interaction of the adenoviral IVa2 protein with a truncated viral DNA packaging sequence.
Adenoviral (Ad) infection typically poses little health risk for immunosufficient individuals. However, for immunocompromised individuals, such as AIDS patients and organ transplant recipients, especially pediatric heart transplant recipients, Ad infection is common and can be lethal. Ad DNA packaging is the process whereby the Ad genome becomes encapsulated by the viral capsid. Specific packaging is dependent upon the packaging sequence (PS), which is composed of seven repeated elements called A repeats. The Ad protein, IVa2, which is required for viral DNA packaging, has been shown to bind specifically to synthetic DNA probes containing A repeats I and II, however, the molecular details of this interaction have not been investigated. In this work we have studied the binding of a truncated form of the IVa2 protein, that has previously been shown to be sufficient for virus viability, to a DNA probe containing A repeats I and II. We find that the IVa2 protein exists as a monomer in solution, and that a single IVa2 monomer binds to this DNA with high affinity (K(d)< approximately 10 nM), and moderate specificity, and that the trIVa2 protein interacts in a fundamentally different way with DNA containing A repeats than it does with non-specific DNA. We also find that at elevated IVa2 concentrations, additional binding, beyond the singly ligated complex, is observed. When this reaction is modeled as representing the binding of a second IVa2 monomer to the singly ligated complex, the K(d) is 1.4+/-0.7 microM, implying a large degree of negative cooperativity exists for placing two IVa2 monomers on a DNA with adjacent A repeats.
Yang
Teng-Chieh
TC
University of Colorado Denver, Department of Pharmaceutical Sciences, School of Pharmacy C238-P15, P.O Box 6511, Aurora, CO 80045, USA.
Yang
Qin
Q
Maluf
Nasib Karl
NK
D000256:Adenoviridae / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D042003:DNA Packaging; D004279:DNA, Viral / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D008954:Models, Biological; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship; D014461:Ultracentrifugation; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism*
D004279:DNA, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; C056232:iva2 protein, adenovirus