Mammary tumor virus DNA contains sequences required for its hormone-regulated transcription

Glucocorticoids regulate the rate of transcription of integrated murine mammary tumor virus (MTV) genes in most clones of MTV-infected rat hepatoma tissue culture (HTC) cells. To determine whether hormonal regulation is mediated from flanking cellular sequences or rather from within the viral DNA, we analyzed the relative rates of transcription of MTV and adjacent HTC sequences in two lines of infected HTC cells, J2.15 and J2.17, each of which contains a single insertion of MTV DNA. In addition, we measured in uninfected HTC cells the transcriptional rates of the corresponding "preinsertion fragments," which may be viewed as the equivalent sequences bearing deletions of MTV DNA; the two genomic segments into which integration occurred in the two lines are unrelated. Previous work showed that glucocorticoids induce the accumulation of MTV RNA in J2.17, whereas viral transcripts are not detected in J2.15, RNA pulse-labeling experiments indicate that glucocorticoids stimulate that rate of MTV gene transcription in J2.17 but not in J2.15; in contrast, no labeled RNA hybridizing to flanking sequences was detected either in the uninfected or in the infected cells. We conclude that the host site of integration cells. We conclude that the host site of integration of MTV need not be transcriptionally active or hormonally responsive to permit viral gene expression. Furthermore, these rate measurements indicate that glucocorticoid-stimulated MTV RNA synthesis in J2.17 does not reflect readthrough transcription from a regulated cellular promoter. This notion is independently supported by transcript mapping experiments showing that the 5' terminus of nuclear MTV RNA is at a site on MTV DNA approximately 1.2 kb downstream from the host-viral junction. Thus our data are consistent with the presence of a hormone-responsive element within the provirus.