Immunological studies of a 21 kDa cellular component efficiently incorporated into rabies virion grown in a BHK-21 cell culture.
To investigate cellular components incorporated into the rabies virion, monoclonal antibodies (MAbs) were screened based on their reactivity with additional virion components. Two of the MAbs we prepared recognized a virion-associated 21 kDa polypeptide (referred to as VAP21) from a BHK-21 cell. Since the MAbs precipitated the rabies virion and trypsin digestion eliminated the VAP21 antigen from the virion but alkaline treatment (pH 11) did not, VAP21 seems to be anchored into the viral envelope and exposed on the virion surface. Although quantitative immunoblot analyses indicated an apparently increased concentration of VAP21 in the virion, the ratio of the content of VAP21 to that of viral glycoprotein (G) was several times decreased as compared to the ratio of those in the cell. These data suggest that sorting of VAP21 occurs during the viral budding process on the cell but that it might be inefficient, probably due to a more intimate association of VAP21 with the viral envelope proteins. This assumption seems to be consistent with the results of immunofluorescence studies; that is, VAP21 displayed colocalized distribution with viral envelope antigens in the cell. From these results, it is suggested that VAP21 closely associates with the viral envelope proteins in the cell, and this association might cause passive but relatively efficient incorporation of VAP21 into the virion.
Sagara
J
J
Department of Molecular Microbiology, Graduate School of Pharmaceutical Sciences, Kyoto University, Japan.
Tochikura
T S
TS
Yamamoto
T
T
Tsukita
S
S
Tsukita
S
S
Kawai
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology; D051379:Mice; D011820:Rabies virus / Q000737:chemistry*; D051381:Rats; D014357:Trypsin / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000276:immunology; D014763:Viral Matrix Proteins / Q000032:analysis / Q000276:immunology; D014771:Virion / Q000737:chemistry* / Q000302:isolation & purification / Q000502:physiology; D019065:Virus Assembly; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; D010450:Endopeptidases; D014357:Trypsin