title: Monoclonal antibodies against regions topologically surrounding the homodimeric beta-barrel interface of Epstein-Barr virus nuclear antigen-1.
abstract: Epstein-Barr virus (EBV) nuclear antigen-1 (EBNA-1) is essential for maintenance of EBV latency. Four mouse monoclonal antibodies (mAbs) against the part of the EBNA-1 sequence (amino acids 451-641) containing the domain that forms a homodimeric eight-stranded beta-barrel were generated and characterized, examined for immunocytochemical staining, immunoblotting and isoelectric focusing of EBNA-1 proteins, and used to examine interactions between EBNA-1 polypeptides by far-Western blot assays. Far-Western blot analyses using the mAbs suggest that both the beta-strand (aa 593-604) and alpha helix (aa 568-582) are essential for EBNA-1 dimerization, consistent with yeast two-hybrid studies of mutant EBNA-1 polypeptides. These mAbs should be useful for studies on the structure and function of EBNA-1 proteins.
authors.affiliation: Herpesvirus Laboratory Department of Virology I, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Eda
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Ishii
authors.forename: Maya
authors.initials: M
authors.lastname: Obayashi
authors.forename: Shizuko
authors.initials: S
authors.lastname: Harada
authors.forename: Sayuri
authors.initials: S
authors.lastname: Ito
authors.forename: Tomomichi
authors.initials: T
authors.lastname: Fujita
authors.forename: Masato
authors.initials: M
authors.lastname: Ikeda
authors.forename: Shuichi
authors.initials: S
authors.lastname: Kusano
authors.forename: Ryo
authors.initials: R
authors.lastname: Kitamura
authors.forename: Chieko
authors.initials: C
authors.lastname: Suzuki
authors.forename: Takahiko
authors.initials: T
authors.lastname: Hara
authors.forename: Motoo
authors.initials: M
authors.lastname: Watanabe
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Satoh
authors.forename: Keisuke
authors.initials: K
authors.lastname: Sugihara
authors.forename: Kazuo
authors.initials: K
authors.lastname: Yanagi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D015153:Blotting, Western; D019281:Dimerization; D019309:Epstein-Barr Virus Nuclear Antigens / Q000276:immunology*; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry; D007525:Isoelectric Focusing; D051379:Mice; D017434:Protein Structure, Tertiary
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D019309:Epstein-Barr Virus Nuclear Antigens; C417029:EBV-encoded nuclear antigen 1