EBNA1 distorts oriP, the Epstein-Barr virus latent replication origin.
The Epstein-Barr virus nuclear antigen 1 (EBNA1) protein binds and activates the latent replication origin (oriP) of the Epstein-Barr virus. We have been studying EBNA1 to determine how it activates replication at oriP. Here we demonstrate that upon binding of EBNA1 to oriP, two thymine residues become reactive to potassium permanganate (KMnO4), indicating a helical distortion at these sites. The KMnO4-reactive thymines are 64 bp apart in the region of dyad symmetry of oriP. Dimethyl sulfate protection studies indicated that EBNA1 binds on the opposite face of the helix from the reactive thymines. The nature of the helical distortion induced by EBNA1 and its possible significance to the initiation of replication are discussed.
Frappier
L
L
Howard Hughes Medical Institute, Microbiology Department, Cornell University Medical College, New York, New York 10021.
O'Donnell
M
M
D000956:Antigens, Viral / Q000502:physiology*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000737:chemistry*; D019309:Epstein-Barr Virus Nuclear Antigens; D004854:Herpesvirus 4, Human / Q000235:genetics*; D066298:In Vitro Techniques; D008745:Methylation; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011196:Potassium Permanganate / Q000737:chemistry; D011994:Recombinant Proteins; D012045:Regulatory Sequences, Nucleic Acid
D000956:Antigens, Viral; D004268:DNA-Binding Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D011196:Potassium Permanganate; D004247:DNA