Visualization of antigens attached to cytoskeletal framework in animal cells: colocalization of simian virus 40 Vp1 polypeptide and actin in TC7 cells.
Actin and the simian virus 40 viral structural polypeptide Vp1 are observed to be present on cytoskeletal fibers of virus-infected TC7 cells, when these antigens in detergent-extracted whole cell mounts were labeled by specific antibodies and colloidal gold particles coated with a second antibody. In both cases, actin and Vp1 were found associated with fibers and fiber-associated electron-dense materials. Patches or clusters of colloidal gold particles denoting the presence of either Vp1 or actin were found on fibers uniformly distributed throughout the cytoplasm. By using simultaneous decoration of the two antigens with colloidal gold particles of different diameters, it was shown that the majority of Vp1 appears attached to cytoskeletal fibers in association with cellular actin. When Vp1 and actin were decorated with Imposil and ferritin simultaneously in infected cells that were fixed first and then permeabilized with saponin, both labels were found in the same spatial domain of the cell cytoplasm. Thus, the colocalization of Vp1 and actin on the cytoskeletal framework seems to reflect their actual state in the living cells. The electron-dense material to which colloidal gold particles localize in our cytoskeletal preparations may be the remnants of subcellular structures with which actin and Vp1 are both associated in intact cells.
D000199:Actins / Q000032:analysis*; D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis*; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology / Q000648:ultrastructure; D003102:Colloids; D005455:Fluorescent Antibody Technique; D006046:Gold; D008854:Microscopy, Electron; D013539:Simian virus 40 / Q000276:immunology*; D014764:Viral Proteins / Q000032:analysis*
D000199:Actins; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D003102:Colloids; D014764:Viral Proteins; D006046:Gold