Mapping of antigenic domains of Sendai virus nucleocapsid protein expressed in Escherichia coli.
Several nonoverlapping epitopes were mapped on the primary sequence of the Sendai virus NP protein. After a complete cDNA clone of the Sendai virus NP gene was expressed in Escherichia coli, deletion constructs were used to generate a series of overlapping NP fragments deleted at their C termini. Immunoblot analyses with 11 monoclonal antibodies identified four antigenic sites. All of these sites resided in the C-terminal half of NP and were also the only sites detected with a polyclonal serum. These findings confirm and extend the evidence that the C terminus of the NP protein represents the domain exposed on the surface of the nucleocapsid. One of the monoclonal antibodies reacted with a site, comprising only 6 amino acids, lying with a hinge between an alpha-helix and a beta-strand in the predicted secondary structure of NP. Since this antibody is a potent inhibitor of in vitro viral RNA synthesis (K. L. Deshpande and A. Portner, Virology 139:32-42, 1984), the epitope may be critical to the flexibility of the NP molecule that makes the RNA template accessible during RNA synthesis.
Gill
D S
DS
Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101-0318.
Takai
S
S
Portner
A
A
Kingsbury
D W
DW
D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis*; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D002872:Chromosome Deletion; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics; D000939:Epitopes / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D015151:Immunoblotting; D010222:Parainfluenza Virus 1, Human / Q000235:genetics / Q000276:immunology*; D010957:Plasmids; D015183:Restriction Mapping; D014758:Viral Core Proteins / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D004279:DNA, Viral; D000939:Epitopes; D014758:Viral Core Proteins; D004262:DNA Restriction Enzymes