title: Two domains of the epstein-barr virus origin DNA-binding protein, EBNA1, orchestrate sequence-specific DNA binding.
abstract: The EBNA1 (for Epstein-Barr nuclear antigen 1) protein of Epstein-Barr virus governs the replication and partitioning of the viral genomes during latent infection by binding to specific recognition sites in the viral origin of DNA replication. The crystal structure of the DNA binding portion of the EBNA1 protein revealed that this region comprises two structural motifs; a core domain, which mediates protein dimerization and is structurally homologous to the DNA binding domain of the papillomavirus E2 protein, and a flanking domain, which mediated all the observed sequence-specific contacts. To test the possibility that the EBNA1 core domain plays a role in sequence-specific DNA binding not revealed in the crystal structure, we examined the effects of point mutations in potential hydrogen bond donors located in an alpha-helix of the EBNA1 core domain whose structural homologue in E2 mediates sequence-specific DNA binding. We show that these mutations severely reduce the affinity of EBNA1 for its recognition site, and that the core domain, when expressed in the absence of the flanking domain, has sequence-specific DNA binding activity. Flanking domain residues were also found to contribute to the DNA binding activity of EBNA1. Thus, both the core and flanking domains of EBNA1 play direct roles in DNA recognition.
authors.affiliation: Department of Medical Genetics and Microbiology, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors.forename: J
authors.initials: J
authors.lastname: Cruickshank
authors.forename: K
authors.initials: K
authors.lastname: Shire
authors.forename: A R
authors.initials: AR
authors.lastname: Davidson
authors.forename: A M
authors.initials: AM
authors.lastname: Edwards
authors.forename: L
authors.initials: L
authors.lastname: Frappier
mesh_terms: D001665:Binding Sites / Q000235:genetics; D004247:DNA / Q000378:metabolism*; D019309:Epstein-Barr Virus Nuclear Antigens / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D004854:Herpesvirus 4, Human / Q000502:physiology; D011485:Protein Binding; D011487:Protein Conformation; D014779:Virus Replication
chemical_list: D019309:Epstein-Barr Virus Nuclear Antigens; D004247:DNA