Monoclonal antibodies against regions topologically surrounding the homodimeric beta-barrel interface of Epstein-Barr virus nuclear antigen-1.
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.
Eda
Hiroyuki
H
Herpesvirus Laboratory Department of Virology I, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan.
Ishii
Yasuyuki
Y
Obayashi
Maya
M
Harada
Shizuko
S
Ito
Sayuri
S
Fujita
Tomomichi
T
Ikeda
Masato
M
Kusano
Shuichi
S
Kitamura
Ryo
R
Suzuki
Chieko
C
Hara
Takahiko
T
Watanabe
Motoo
M
Satoh
Hiroshi
H
Sugihara
Keisuke
K
Yanagi
Kazuo
K
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
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D019309:Epstein-Barr Virus Nuclear Antigens; C417029:EBV-encoded nuclear antigen 1