Binding of SV40 a protein to the BK virus origin of DNA replication

Abstract

The SV40 A protein (T antigen) binds to the putative origin of replication of the related BK virus (BKV). Protection studies with dimethyl sulfate identify multiple pentanucleotide contact sites in three distinct binding regions. In addition to the consensus family of recognition sequences, 5'-(G greater than T)(A greater than G)GGC-3', previously identified in the origin of SV40, BKV contains a contact 5'-AAGGC-3' site. As in the case of SV40, each binding region contains recognition sequences in different arrangements. The high affinity region I contains two sites arranged as direct repetitions covering a span of 17 base pairs (bp). In the intermediate affinity region II, four pentanucleotides are oriented as inverted repetitions with a span of 23 bp. Low affinity region III has a single contact site that can direct binding of the A protein. These different arrangements of DNA contact sites determine different patterns of protein binding and DNase protection in the three regions. The findings support a model of A protein binding to DNA previously proposed for the SV40 origin and establish a basis for future studies on regulatory phenomena at the BKV origin.