Extra mouse mammary tumor proviruses in DBA/2 mouse lymphomas acquire a selective advantage in lymphocytes by alteration in the U3 region of the long terminal repeat

Abstract

We determined the nucleotide sequences of the long terminal repeats (LTRs) from mouse mammary tumor virus (MMTV) proviruses acquired in two DBA/2 mouse lymphoma cell lines, MLA and DL-8. Proviruses from MLA contained a 352-base-pair deletion from nucleotides 669 to 1020 in the U3 region of the LTR, whereas the LTR alteration of the DL-8 provirus involved both a similar 360-base-pair deletion and generation of a tandem repeat region consisting of sequences of flanking deletions. To assess the function of the rearranged LTRs, we constructed plasmids in which normal and rearranged LTRs drove the reporter chloramphenicol acetyltransferase gene and transfected them into T-cell lines (Jurkat, Molt-3, and DL-8) and the mammary tumor cell line T47D. Both rearranged LTRs were transcriptionally active, but normal LTRs were not active in either the presence or absence of glucocorticoids in all T-cell lines. In T47D cells, however, the MLA provirus LTR showed the same glucocorticoid- or progestin-dependent transcriptional activity as did normal LTRs. The DL-8 provirus LTR acquired a novel enhancer(s) by rearrangement and thus had a high basal transcriptional activity in T47D cells. The results of chloramphenicol acetyltransferase assays using plasmids with various chimeric MMTV LTRs revealed that the rearranged LTRs had lost their negative regulatory element and contained an enhancer element that was highly homologous to the enhancer A element of polyomavirus (from nucleotides 525 to 558). GR but not C3H mouse MMTV contained this enhancer. These results elucidate some of the molecular mechanisms involved in the selection of mutant MMTVs with rearranged LTRs in lymphoma cells.