The rabies virion-associated 100-kDa polypeptide (VAP100) is a host-derived minor component of the viral envelope.
We investigated a minor polypeptide component of 100-kDa detected in the rabies virion (referred to as VAP100) by using a monoclonal antibody (mAb), #16743, which was shown to recognize the SDS-denatured VAP100 antigen by immunoblot analyses. Although the VAP100 antigen was hardly detectable in the cell by usual immunoblot methods with this mAb, we could detect the antigen by a luminescent immunoblot method as well as by immunoprecipitation from the metabolically radiolabeled cell lysates and virions. Fluorescent antibody (FA) staining with mAb #16743 detected the uniformly distributed antigen on the formalin-fixed normal BHK-21 cells, while slight accumulation of the antigen was also seen in the Golgi area when the cells were permeabilized by treatment with Triton X-100 after fixation. Rabies virus infection induced alteration of the behavior of VAP100 to show a spotted distribution pattern in virus-infected cells. Double FA staining with mAb #16743 and rabbit antibody against the rabies virus envelope antigen demonstrated colocalized distribution of the viral envelope antigens and VAP100 in the cell. From these results, we think that VAP100 is a membrane-associated component of the cell, and its colocalized distribution with the viral envelope antigens in the cell implicates an intimate association of the VAP100 with viral envelope protein(s) and a reflection of possible involvement in the efficient incorporation of VAP100 into the virion.
Xiao
S
S
Department of Molecular Microbiology, Graduate School of Pharmaceutical Sciences, Kyoto University, Japan.
Komiya
K
K
Tochikura
T S
TS
Sagara
J
J
Tsukita
S
S
Kawai
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D019556:COS Cells / Q000821:virology; D002460:Cell Line / Q000821:virology*; D006367:HeLa Cells / Q000821:virology; D006801:Humans; D019169:Jurkat Cells / Q000821:virology; D008970:Molecular Weight; D010455:Peptides / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D011820:Rabies virus / Q000737:chemistry*; D014759:Viral Envelope Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D014771:Virion / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D010455:Peptides; D014759:Viral Envelope Proteins