Permissive infections of BHK cells and nonpermissive infections of Raji cells were probed for the accumulation of vesicular stomatitis virus intracellular RNAs. In Raji cells, the onset of vesicular stomatitis virus transcription and replication was delayed when compared to BHK cells, and the accumulation of plus and minus sense leader RNAs was significantly reduced. In contrast, full length plus and minus strand replicative RNAs accumulated in Raji cells to levels approximately equivalent to those in BHK cells. In both cell types, approximately four times as many minus strands as plus strands were detected late in the infections. At 16 h postinfection, 12% of the total genomic RNA synthesized in BHK cells was packaged and released whereas only 0.8% was released from Raji cells. In addition, of those particles released by Raji cells, only 1% were infectious whereas 77% of those released by BHK cells were infectious. The virions released from both cell types contained similar amounts of the five viral proteins, however. Analysis of virions from Raji cells revealed a faster electrophoretic mobility of the glycoprotein than the glycoprotein in virions released from BHK cells. These results suggest that Raji cells may be restricted in their ability to support a complete infection at the level of virus assembly rather than at the level of RNA replication.