title: Comparative inhibition of rabbit globin mRNA translation by modified antisense oligodeoxynucleotides.
abstract: We have studied the translation of rabbit globin mRNA in cell free systems (reticulocyte lysate and wheat germ extract) and in microinjected Xenopus oocytes in the presence of anti-sense oligodeoxynucleotides. Results obtained with the unmodified all-oxygen compounds were compared with those obtained when phosphorothioate or alpha-DNA was used. In the wheat germ system a 17-mer sequence targeted to the coding region of beta-globin mRNA was specifically inhibitory when either the unmodified phosphodiester oligonucleotide or its phosphorothioate analogue were used. In contrast no effect was observed with the alpha-oligomer. These results were ascribed to the fact that phosphorothioate oligomers elicit an RNase-H activity comparable to the all-oxygen congeners, while alpha-DNA/mRNA hybrids were a poor substrate. Microinjected Xenopus oocytes followed a similar pattern. The phosphorothioate oligomer was more efficient to prevent translation than the unmodified 17-mer. Inhibition of beta-globin synthesis was observed in the nanomolar concentration range. This result can be ascribed to the nuclease resistance of phosphorothioates as compared to natural phosphodiester linkages, alpha-oligomers were devoid of any inhibitory effect up to 30 microM. Phosphorothioate oligodeoxyribonucleotides were shown to be non-specific inhibitors of protein translation, at concentrations in the micromolar range, in both cell-free systems and oocytes. Non-specific inhibition of translation was dependent on the length of the phosphorothioate oligomer. These non-specific effects were not observed with the unmodified or the alpha-oligonucleotides.
authors: Laboratoire de Biophysique, INSERM U201, Paris, France.
authors: C
authors: C
authors: Cazenave
authors: C A
authors: CA
authors: Stein
authors: N
authors: N
authors: Loreau
authors: N T
authors: NT
authors: Thuong
authors: L M
authors: LM
authors: Neckers
authors: C
authors: C
authors: Subasinghe
authors: C
authors: C
authors: Hélène
authors: J S
authors: JS
authors: Cohen
authors: J J
authors: JJ
authors: Toulmé
mesh_terms: D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D002474:Cell-Free System; D005914:Globins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000138:chemical synthesis / Q000494:pharmacology; D009865:Oocytes / Q000378:metabolism; D014176:Protein Biosynthesis; D012313:RNA; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D013816:Thermodynamics; D014908:Triticum / Q000235:genetics; D014981:Xenopus / Q000235:genetics
chemical_list: D015345:Oligonucleotide Probes; D016372:RNA, Antisense; D012333:RNA, Messenger; D012313:RNA; D005914:Globins; D012260:Ribonucleases