Mouse polyomavirus enters host cells internalized, similar to simian virus 40 (SV40), in smooth monopinocytic vesicles, the movement of which is associated with transient actin disorganization. The major
capsid protein (VP1) of the incoming polyomavirus accumulates on membranes around the cell nucleus. Here we show that unlike SV40, mouse
polyomavirus infection is not substantially inhibited by
brefeldin A, and colocalization of VP1 with
beta-COP during early stages of
polyomavirus infection in mouse fibroblasts was observed only rarely. Thus, these viruses obviously use different traffic routes from the plasma membrane toward the cell nucleus. At approximately 3 h postinfection, a part of VP1 colocalized with the endoplasmic reticulum marker BiP, and a subpopulation of virus was found in perinuclear areas associated with Rab11
GTPase and colocalized with
transferrin, a marker of recycling endosomes. Earlier postinfection, a minor subpopulation of virions was found to be associated with Rab5, known to be connected with early endosomes, but the cell entry of virus was slower than that of
transferrin or
cholera toxin B-fragment. Neither Rab7, a marker of late endosomes, nor LAMP-2 lysosomal
glycoprotein was found to colocalize with polyomavirus. In situ hybridization with polyomavirus genome-specific
fluorescent probes clearly demonstrated that, regardless of the multiplicity of
infection, only a few virions delivered their genomic
DNA into the cell nucleus, while the majority of viral genomes (and VP1) moved back from the proximity of the nucleus to the cytosol, apparently for their degradation.