@article{mbs:/content/journal/jgv/10.1099/0022-1317-81-1-119, author = "Goto, Hideo and Minamoto, Nobuyuki and Ito, Hiroshi and Ito, Naoto and Sugiyama, Makoto and Kinjo, Toshio and Kawai, Akihiko", title = "Mapping of epitopes and structural analysis of antigenic sites in the nucleoprotein of rabies virus", journal= "Journal of General Virology", year = "2000", volume = "81", number = "1", pages = "119-127", doi = "https://doi.org/10.1099/0022-1317-81-1-119", url = "https://www.microbiologyresearch.org/content/journal/jgv/10.1099/0022-1317-81-1-119", publisher = "Microbiology Society", issn = "1465-2099", type = "Journal Article", abstract = "Linear epitopes on the rabies virus nucleoprotein (N) recognized by six MAbs raised against antigenic sites I (MAbs 6-4, 12-2 and 13-27) and IV (MAbs 6-9, 7-12 and 8-1) were investigated. Based on our previous studies on sites I and IV, 24 consecutively overlapping octapeptides and N- and C-terminal-deleted mutant N proteins were prepared. Results showed that all three site I epitopes studied and two site IV epitopes (for MAbs 8-1 and 6-9) mapped to aa 358–367, and that the other site IV epitope of MAb 7-12 mapped to aa 375–383. Tests using chimeric and truncated proteins showed that MAb 8-1 also requires the N-terminal sequence of the N protein to recognize its binding region more efficiently. Immunofluorescence studies demonstrated that all three site I-specific MAbs and one site IV-specific MAb (7-12) stained the N antigen that was diffusely distributed in the whole cytoplasm; the other two site IV-specific MAbs (6-9 and 8-1) detected only the N antigen in the cytoplasmic inclusion bodies (CIB). An antigenic site II-specific MAb (6-17) also detected CIB-associated N antigen alone. Furthermore, the level of diffuse N antigens decreased after treatment of infected cells with cycloheximide. These results suggest that epitopes at site I are expressed on the immature form of the N protein, but epitope structures of site IV MAbs 6-9 and 8-1 are created and/or exposed only after maturation of the N protein.", }