title: Transcription from a spleen necrosis virus 5' long terminal repeat is suppressed in mouse cells.
abstract: To determine the block(s) to spleen necrosis virus (SNV) replication in mouse cells, we studied the expression of a dominant selectable marker, neo, or a gene whose product is easily assayed, the chloramphenicol acetyltransferase (cat) gene, in SNV-derived and murine leukemia virus-derived vectors. Using transient (CAT) and stable (Neor phenotype) transfection assays, we showed that the SNV promoter was used in mouse cells only when the 3' SNV long terminal repeat (LTR) was absent. Infection of mouse cells with recombinant SNV viruses was 1% as efficient as infection of permissive dog (D17) cells. The SNV proviruses in mouse cells appeared normal by Southern blot analysis, indicating that their integration probably occurred by normal mechanisms. S1 nuclease analyses of Neor mouse cell clones, each harboring a single recombinant SNV provirus, showed that the selected (internal) promoter was active, but that the 5' SNV LTR promoter was not. However, in the rare (less than 10(-6)) Neor colonies in which expression of the 5' LTR was selected, both promoters were active. Thus, the block to SNV infection of mouse cells is at least at two levels; one is a 100-fold-decreased efficiency at some step(s) up to and including integration, and the other is at transcription.
mesh_terms: D000818:Animals; D002417:Cattle; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D005347:Fibroblasts; D005814:Genes, Viral; D005822:Genetic Vectors; D009052:Leukemia Virus, Murine / Q000235:genetics; D051379:Mice; D010957:Plasmids; D012313:RNA / Q000235:genetics; D001356:Reticuloendotheliosis virus / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics*; D013489:Suppression, Genetic; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D012313:RNA