Epitopes and nuclear localization analyses of canine distemper virus nucleocapsid protein by expression of its deletion mutants.
A series of nucleocapsid protein (NP)-deleted genes of the Onderstepoort strain was constructed in order to locate antigenic regions of the NP of canine distemper virus. The expression of proteins from 5'-deleted NP genes was examined in COS-7 cells by indirect immunofluorescence assay using three monoclonal antibodies (MAbs), c-5, f-5 and h-6, and a rabbit serum against NP. These MAbs reacted with two regions of NP. Amino acid residues from 1 to 80, and 337-358, were necessary and sufficient for formation of the epitopes identified by MAbs f-5 and h-6, and c-5, respectively. The proteins translated from intact or 3'-deleted genes were found to be localized in the nuclei of COS-7 cells, whereas the proteins from the 5'-deleted genes were mainly detected in the cytoplasm. These results suggested that 80 amino acid residues at the N-terminus are required for transportation of NP into the nucleus.
Yoshida
E
E
Department of Veterinary Microbiology, Graduate School of Agriculture and Life Science, The University of Tokyo, Japan.
Shin
Y S
YS
Iwatsuki
K
K
Gemma
T
T
Miyashita
N
N
Tomonaga
K
K
Hirayama
N
N
Mikami
T
T
Kai
C
C
D000818:Animals; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000737:chemistry; D019556:COS Cells; D002522:Chlorocebus aethiops; D017931:DNA Primers / Q000737:chemistry; D004262:DNA Restriction Enzymes / Q000737:chemistry; D004216:Distemper / Q000821:virology*; D004217:Distemper Virus, Canine / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D000939:Epitopes / Q000235:genetics; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D016296:Mutagenesis; D019590:Nucleocapsid Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011233:Precipitin Tests / Q000662:veterinary; D014162:Transfection
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D017931:DNA Primers; D000939:Epitopes; D019590:Nucleocapsid Proteins; D004262:DNA Restriction Enzymes