Vaccinia virion surface polypeptide Ag35 expressed from a baculovirus vector is targeted to analogous poxvirus and insect virus components.
Polypeptide Ag35, a major early component of the vaccinia surface, is integrated into the formative viral lipoprotein tegument. To ascertain whether positioning of Ag35 is due to its general affinity for newly assembled viral membranes we created a recombinant A12 vector to express the vaccinia protein. The baculovirus system was chosen because intranuclear virions of this agent are likewise enclosed inside newly formed envelopes. Comparable infections of two insect cell lines established that more abundant synthesis occurred in High Five (H5) than in SF9 cells. We, therefore, used H5 cells for most experiments reported here. Combined analyses by PAGE, Western blotting, and immunocytology, using light and electron microscopy, revealed a dissemination of Ag35 throughout the cell. Higher concentrations were evident at the cell surface, nuclear perimeter, and within intranuclear virogenic stroma. The association with the virogenic stroma was of specific interest with respect to vaccinia development because it showed a similarity in the targeting of Ag35 toward intranuclear DNA-protein foci of baculovirus which are analogous to the vaccinia-specified cytoplasmic "factories." A further remarkable analogy concerns association of Ag35 with intranuclear baculovirus envelopes, revealing a propensity of Ag35 for nascent viral lipoprotein membranes.
Mohandas
A R
AR
Department of Microbiology and Immunology, University of Western Ontario, London, Canada.
Dekaban
G A
GA
Dales
S
S
D000818:Animals; D000954:Antigens, Surface / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001692:Biological Transport; D002467:Cell Nucleus / Q000382:microbiology; D002478:Cells, Cultured; D003593:Cytoplasm / Q000382:microbiology; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D007304:Insect Viruses / Q000254:growth & development / Q000378:metabolism* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D009036:Moths / Q000166:cytology / Q000648:ultrastructure; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011212:Poxviridae / Q000254:growth & development / Q000378:metabolism* / Q000648:ultrastructure; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D014771:Virion / Q000254:growth & development / Q000648:ultrastructure
D000954:Antigens, Surface; D000956:Antigens, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; C080409:vaccinia surface antigen