Binding of oligopyrimidines to the RNA hairpin responsible for the ribosome gag‐pol frameshift in HIV‐1

FEBS Letters

Toulmé, Jean-Jacques¹  Aupeix, Karine¹  Le Tinévez, Réjane¹ 

[1] INSERM U386, IFR Pathologies Infectieuses, Université Victor Segalen, 146 rue Léo Saignat, F. 33076 Bordeaux Cedex, France

关键词: RNA structure;    Antisense oligonucleotide;    Triple helix;   

DOI  :  10.1016/S0014-5793(99)00427-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown