Rabies virus (RABV) causes a fatal zoonotic
encephalitis. Disease symptoms require replication and spread of the virus within neuronal cells; however, in infected animals as well as in cell culture the virus replicates in a broad range of cell types. Here we use a single-cycle RABV and a recombinant
vesicular stomatitis virus (rVSV) in which the
glycoprotein (G) was replaced with that of RABV (rVSV RABV G) to examine RABV uptake into the African green monkey kidney cell line BS-C-1. Combining biochemical studies and real-time spinning-disk confocal fluorescence microscopy, we show that the predominant entry pathway of RABV particles into BS-C-1 cells is
clathrin dependent. Viral particles enter cells in pits with elongated structures and incomplete
clathrin coats which depend upon actin to complete the internalization process. By measuring the time of internalization and the abundance of the
clathrin adaptor protein AP2, we further show that the pits that internalize RABV particles are similar to those that internalize VSV particles. Pharmacological perturbations of
dynamin or of actin polymerization inhibit productive
infection, linking our observations on particle uptake with viral infectivity. This work extends to RABV particles the finding that
clathrin-mediated endocytosis of rhabdoviruses proceeds through incompletely coated pits which depend upon actin.