Evidence for a prokaryotic promoter in the murine mammary tumor virus long terminal repeat.
The long terminal repeat (LTR) of C3H murine mammary tumor virus (MuMTV) is approx. 1.3 kb long. HaeIII digestion of a cloned PstI fragment containing the left-end LTR generated four fragments of sizes 0.56, 0.41, 0.34 and 0.14 kb, one of which (0.41 kb) had a promoter activity in Escherichia coli. This was demonstrated by replacing the bacterial promoter for the neomycin-resistance (NmR) gene in the plasmid pKC56 with the HaeIII fragments. Only the 0.41-kb fragment that contains sequences from the U3 region of the LTR was found to contain a promoter, as shown by the expression of the drug-resistance phenotype in the recombinant plasmid. The strength of this promoter was comparable to or greater than that found with the parental NmR gene promoter. S1 nuclease mapping of the NmR gene transcript indicated that the initiation of this transcript occurred within the 0.41-kb LTR fragment from a site approx. 10 bp upstream from the 3' end. A comparison of the known DNA sequences in the MuMTV LTR with those found in bacterial promoters revealed that a 'Pribnow box', the initiation signal for the prokaryotic promoters, is present in the 0.41-kb LTR fragment upstream from the initiation site. Furthermore, in a recombinant plasmid that contained the complete LTR the same promoter sequences appeared to be involved in the initiation of RNA transcription. The 0.34-kb LTR fragment, which contains sequences derived from the U3 and U5 regions of the LTR, did not possess promoter activity in E. coli. However, it was found to induce deletions of adjacent plasmid DNA sequences. The deletions were specifically initiated from the downstream end of the LTR-fragment insert. The presence of a prokaryotic promoter in the MuMTV LTR, together with the observation that certain LTR sequences can induce deletions, analogous to those caused by transposable elements, in recombinant plasmids suggest that the MuMTV LTR may have evolved from such elements.
D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics*; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000235:genetics; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics*; D009355:Neomycin / Q000494:pharmacology; D009876:Operon; D010957:Plasmids; D012091:Repetitive Sequences, Nucleic Acid; D014158:Transcription, Genetic
D004279:DNA, Viral; D009355:Neomycin