The production of virus and the synthesis of virus-specific RNA has been studied in Fv-1n/n (NIH/3T3, SIM) and Fv-1b/b (BALB/3T3, SIM-R) cell lines after infection with N- or B-tropic MuLV. It was found that virus production, measured by reverse transcriptase activity in the medium, was 70-100 fold lower in cells resistant at the Fv-1 locus than in permissive cells. The virus-specific RNA, detected by hybridization. In RNA excess with complementary DNA, was reduced by approximately 70-100 fold in cytoplasm of resistant cells compared to permissive cells. A reduction of the same magnitude was observed in the levels of virus-specific RNA extracted from nuclei of resistant cells. Our data therefore show that virus-specific RNA levels are reduced in cells nonpermissive at the Fv-1 locus, suggesting that restriction of the Fv-1 gene product occurs at the level of transcription of the viral genome or at a pre-integration step, or, alternatively, that the RNA transcripts are rapidly degraded after their synthesis.