Interaction of Epstein-Barr viral (EBV) origin of replication (oriP) with EBNA-1 and cellular anti-EBNA-1 proteins.
We have previously shown that 12-O-tetradecanoylphorbol-13-acetate (TPA) which activates expression of the latent genome of the Epstein-Barr virus (EBV) in Burkitt lymphoma cells induces the synthesis of two cellular anti-EBNA-1 competitor proteins, anti-EBNA-1.1 and anti-EBNA-1.2. Both anti-EBNA-1 proteins can uncouple the specific binding of the EBNA-1 to the region required for EBV plasmid maintenance (oriP). Here, we show by DNase I footprinting that the binding sites on oriP for the EBNA-1 and the anti-EBNA-1 proteins were indistinguishable. The proteins bound to the 30-bp tandem repeats of the oriP. Glycerol-gradient centrifugation and gel retardation assay revealed that a 60-kDa protein formed the anti-EBNA-1.1-DNA complex and a 40-kDa protein formed the anti-EBNA-1.2-DNA complex.
Wen
L T
LT
Laboratory of Virology, Tampa Bay Research Institute, St. Petersburg, Florida 33716.
Lai
P K
PK
Bradley
G
G
Tanaka
A
A
Nonoyama
M
M
D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D002478:Cells, Cultured; D004279:DNA, Viral / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology*; D003850:Deoxyribonuclease I; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D012091:Repetitive Sequences, Nucleic Acid; D014779:Virus Replication
D000956:Antigens, Viral; D004279:DNA, Viral; D004268:DNA-Binding Proteins; C033305:Epstein-Barr virus early antigen; D003850:Deoxyribonuclease I