Abstract
To better define protein-DNA interactions at a eukaryotic origin, the domain of simian virus 40 (SV40) large T antigen that specifically interacts with the SV40 origin has been purified and its binding to DNA has been characterized. Evidence is presented that the affinity of the purified T antigen DNA-binding domain for the SV40 origin is comparable to that of the full-length T antigen. Furthermore, stable binding of the T antigen DNA- binding domain to the SV40 origin requires pairs of pentanucleotide recognition sites separated by approximately one turn of a DNA double helix and positioned in a head-to-head orientation. Although two pairs of pentanucleotides are present in the SV40 origin, footprinting and band shift experiments indicate that binding is limited to dimer formation on a single pair of pentanucleotides. Finally, it is demonstrated that the T antigen DNA- binding domain interacts poorly with single-stranded DNA.
Original language | English (US) |
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Pages (from-to) | 3972-3985 |
Number of pages | 14 |
Journal | Journal of virology |
Volume | 71 |
Issue number | 5 |
DOIs | |
State | Published - May 1997 |
ASJC Scopus subject areas
- Microbiology
- Immunology
- Insect Science
- Virology