DNA binding of polyomavirus large T-antigen: kinetics of interactions with different types of binding sites.
Polyomavirus large T-antigen binds to GRGGC sites in double-stranded viral DNA, regulating transcription and replication. Using surface plasmon resonance to record interactions of large T-antigen with different types of binding sites, we found that the configuration of recognition motifs influenced both the association and dissociation rates. Particularly, the complex formed at the origin of DNA replication was labile. A comparison of the interactions between large T-antigen and binding sites with one, two and four GRGGC motifs in tandem showed a strong preference for dimer binding, without detectable co-operativity between dimers. Sodium chloride stabilised the complexes, whereas the dissociation increased rapidly by increasing pH above 7.0.
Bondeson
K
K
Department of Medical Immunology and Microbiology, Uppsala University, Biomedical Centre, Sweden.
Rönn
O
O
Magnusson
G
G
D000957:Antigens, Viral, Tumor / Q000378:metabolism*; D001665:Binding Sites; D015374:Biosensing Techniques; D018983:DNA Footprinting; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D011120:Polyomavirus / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D018741:Replication Origin / Q000235:genetics; D012965:Sodium Chloride / Q000494:pharmacology
D000957:Antigens, Viral, Tumor; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D012965:Sodium Chloride