Determination of recognition sites of T4 RNA ligase on the 3'-OH and 5' -P termini of polyribonucleotide chains.
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The availability of a potent inducer of interferon, the synthetic double-stranded RNA (poly I.poly C), prompted us to determine its possible prophylactic and/or therapeutic effect on virus-induced sarcomas of mice. When treatment was begun prior to virus inoculation and repeated on alternate days, the majority of NIH Swiss mice inoculated with Moloney or Friend pseudotypes of Moloney murine sarcoma virus failed to develop tumors. Other experiments suggested that repeated injections initiated even after the establishment of tumor nodules were also effective. Mice injected on the day of birth with poly I.poly C develop high titers of interferon. The evidence favors, but does not establish, the interpretation that the observed tumor inhibition is mediated through the induction of endogenous interferon. These experiments demonstrate that treatment with poly I.poly C is followed by a regression of established murine sarcoma infection in mice.
Noting the scarcity of CpG dinucleotide in total genomic DNA derived from higher organisms and the scarcity of TpA dinucleotide in total genomic DNA derived from most life forms, we examined the distribution of these dinucleotides in sequences derived from functionally distinct types of human DNA, including mitochondrial DNA, intergenic DNA, intron DNA, and DNA destined to be represented in the cytoplasm as mRNA, tRNA, or rRNA. While CpG frequency has fallen to its lowest levels in DNA that is transcriptionally silent, TpA is most stringently excluded in DNA destined to be expressed as mRNA in the cytosol. This observation suggests that the selective pressures leading to the removal of CpG and TpA operate at different levels. With respect to TpA, dinucleotide scarcity may reflect a requirement for mRNA stability and may indicate the action of UpA-selective ribonucleases. We propose that, by reason of its instability, UpA must have been very rare in primordial RNA. Therefore, tRNA with the anticodon for this dinucleotide may have failed to evolve, making UpA the primordial doublet "stop" codon. The modern triplet code has faithfully conserved this arrangement in the two universal stop codons, UAA and UAG.
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We have shown that cytotoxic T cells can be polyclonally activated by a short exposure to complexes of polyadenylic:polyuridylic acid (poly A:U). Activation is optimal at a dose of 100 microgram/ml poly A:U and occurs during a 2 h incubation period in the absence of antigen. Killing of allogeneic, but not syngeneic, target cells can be observed after 12 h in culture and peaks after 21-24 h in the absence of any nonspecific binding ligand. The observed cytotoxicity is mediated by T lymphocytes and dose not require accessory macrophages or DNA synthesis for the activation or expression of effector functions. These results suggest that few requirements exist for the activation of cytotoxic T cells.
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