Inhibition of in vitro and ex vivo translation by a transplatin-modified oligo(2'-O-methylribonucleotide) directed against the HIV-1 gag-pol frameshift signal

• Published in: Nucleic acids research Vol. 28; no. 2; pp. 438 - 445
• Main Authors: Aupeix-Scheidler, K, Chabas, S, Bidou, L, Rousset, J P, Leng, M, Toulmé, J J
• Format: Journal Article
• Language: English
• Published: England Oxford Publishing Limited (England) 15.01.2000; Oxford University Press
• Subjects: 3T3 Cells; Animals; Base Sequence; Cisplatin - chemistry; Frameshifting, Ribosomal; Fusion Proteins, gag-pol - genetics; HIV-1 - genetics; Mice; Molecular Sequence Data; Oligoribonucleotides - chemistry; Oligoribonucleotides - pharmacology; Protein Biosynthesis - drug effects; RNA, Viral - genetics; Transcription, Genetic - drug effects
• ISSN: 1362-4962; 0305-1048; 1362-4962
• DOI: 10.1093/nar/28.2.438

A 2'-O-methylribooligonucleotide containing a G1.U.G3 triad modified by trans-diamminedichloro-platinum(II) was targeted to the RNA region responsible for the gag-pol frameshifting during translation of the HIV-1 mRNA. The binding of the platinated oligonucleotide to its target RNA induced a rearrangement of the (G1, G3)-intrastrand crosslink, leading to the formation of an intermolecular oligonucleotide-RNA G-A crosslink. This resulted in the selective arrest of translation of a luciferase gene placed downstream of the HIV-1 frameshift signal both in a cell-free extract (rabbit reticulocyte lysate) and in RNA-transfected cells. A specific inhibition of luciferase activity was still observed when the oligonucleotide-RNA complex was not pre-formed prior to either translation or transfection. Moreover, a selective inhibition was also observed when the oligonucleotide and the plasmid DNA encoding the luciferase and bearing the RNA gag- pol frameshifting signal were co-transfected in NIH 3T3 cultured cells. Therefore the intra-strand-->interstrand conversion of the platinum crosslink kinetically competes with the translation machinery and blocks the polypeptide elongation. These transplatin-modified oligonucleotides which operate within a live cell on a 'real-time' basis and do not need an external triggering signal constitute a promising new class of selective reactive probes.