Phorbol ester-inducible T-cell-specific expression of variant mouse mammary tumor virus long terminal repeats.
Acquired proviruses of mouse mammary tumor virus (MMTV) in T-cell leukemias of male GR mice have rearrangements in the U3 region of their long terminal repeats (LTR). In contrast to the endogenous nonrearranged MMTV proviruses, these mutated copies are highly expressed in leukemic T cells. To investigate whether the sequence alterations in the LTR are responsible for the high expression of rearranged MMTV proviruses, we made constructs in which normal and variant LTRs drive the bacterial reporter gene chloramphenicol acetyltransferase (CAT). Two different rearranged LTRs were used, one containing a 420-base-pair (bp) deletion (L13) and another carrying a 456-bp deletion plus an 82-bp insertion (L42). These constructs were transfected into murine (GRSL) and human (MOLT-4) T-cell lines that either had or had not been treated with phorbol ester (12-O-tetradecanoylphorbol-13-acetate [TPA]). In GRSL cells, the L13-LTR-CAT construct showed transcriptional activity that was further enhanced by TPA. In MOLT-4 cells, both variant LTRs were active, but only after stimulation with TPA. In contrast, normal(N)-LTR-CAT constructs were not expressed, irrespective of TPA addition. In XC rat fibrosarcoma cells, neither normal nor variant LTRs gave rise to detectable CAT activity, either in the presence or in the absence of TPA, but dexamethasone strongly stimulated CAT activity driven by N and L42 LTRs. The L13 LTR was considerably less active, probably caused by the deletion of the distal part of the glucocorticoid responsive element. We conclude that the LTR rearrangements generate TPA responsiveness and contribute to T-cell-specific expression of MMTV variants.
Theunissen
H J
HJ
Division of Molecular Biology, Antoni van Leeuwenhoekhuis, Netherlands Cancer Institute, Amsterdam.
Paardekooper
M
M
Maduro
L J
LJ
Michalides
R J
RJ
Nusse
R
R
D000818:Animals; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D005786:Gene Expression Regulation; D015321:Gene Rearrangement; D006801:Humans; D015458:Leukemia, T-Cell / Q000382:microbiology*; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics*; D051379:Mice; D011533:Proviruses / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
D004279:DNA, Viral; D013755:Tetradecanoylphorbol Acetate