RT Journal Article SR Electronic(1) A1 Goto, Hideo A1 Minamoto, Nobuyuki A1 Ito, Hiroshi A1 Ito, Naoto A1 Sugiyama, Makoto A1 Kinjo, Toshio A1 Kawai, Akihiko YR 2000 T1 Mapping of epitopes and structural analysis of antigenic sites in the nucleoprotein of rabies virus JF Journal of General Virology, VO 81 IS 1 SP 119 OP 127 DO https://doi.org/10.1099/0022-1317-81-1-119 PB Microbiology Society, SN 1465-2099, AB 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., UL https://www.microbiologyresearch.org/content/journal/jgv/10.1099/0022-1317-81-1-119