title: Purification of the simian virus 40 (SV40) T antigen DNA-binding domain and characterization of its interactions with the SV40 origin.
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.
authors.affiliation: Department of Biochemistry, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
authors.forename: W S
authors.initials: WS
authors.lastname: Joo
authors.forename: X
authors.initials: X
authors.lastname: Luo
authors.forename: D
authors.initials: D
authors.lastname: Denis
authors.forename: H Y
authors.initials: HY
authors.lastname: Kim
authors.forename: G J
authors.initials: GJ
authors.lastname: Rainey
authors.forename: C
authors.initials: C
authors.lastname: Jones
authors.forename: K R
authors.initials: KR
authors.lastname: Sreekumar
authors.forename: P A
authors.initials: PA
authors.lastname: Bullock
mesh_terms: D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification / Q000378:metabolism*; D013539:Simian virus 40 / Q000276:immunology*
chemical_list: D000952:Antigens, Polyomavirus Transforming; D010446:Peptide Fragments; D004247:DNA