Binding of oligopyrimidines to the RNA hairpin responsible for the ribosome gag-pol frameshift in HIV-1.
The 12 bp stem of the RNA hairpin responsible for the gag-pol frameshifting of the ribosomes during translation of the polycistronic HIV-1 mRNA has a pyrimidine-rich 5' strand and, consequently, a purine-rich 3' strand. Electrophoretic mobility shift assays have shown that DNA oligopyrimidines, 12 and 20 nucleotides long (but not oligopurines or G,T-containing oligomers), designed to form triplexes actually bind to the double-stranded RNA target. RNase V1 footprinting studies have confirmed the interaction between the hairpin stem and the RNA and 2'-O-methyl oligoribonucleotide analogues of the 12-mer oligodeoxypyrimidine as well as 5 propynyl,cytosine, containing the 12-mer oligodeoxypyrimidine, bind more strongly to the RNA target than the unmodified parent DNA oligomer. The complexes formed by the RNA hairpin and either the 12-mer oligodeoxypyrimidine or the 20-mer oligopyrimidine are stable at a neutral pH and in the absence of Mg2+ but blocked neither the reverse transcription nor cell-free translation of a RNA template in which the gag-pol frameshifting hairpin was inserted at the 5' end of the luciferase open reading frame.
Aupeix
K
K
INSERM U386, IFR Pathologies Infectieuses, Université Victor Segalen, Bordeaux, France.
Le Tinévez
R
R
Toulmé
J J
JJ
D001681:Biological Assay; D018983:DNA Footprinting; D016368:Frameshift Mutation; D015685:Fusion Proteins, gag-pol / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D011743:Pyrimidines / Q000378:metabolism*; D012330:RNA, Double-Stranded / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D012270:Ribosomes; D012965:Sodium Chloride
D015685:Fusion Proteins, gag-pol; D009838:Oligodeoxyribonucleotides; D011743:Pyrimidines; D012330:RNA, Double-Stranded; D012367:RNA, Viral; D012965:Sodium Chloride