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PMID: 2472605 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Comparative inhibition of rabbit globin mRNA translation by modified antisense oligodeoxynucleotides.

Nucleic acids research ·Vol. 17 ·No. 11 ·1989-06-12 ·Pages 4255-73

Cazenave C, Stein CA, Loreau N, Thuong NT, Neckers LM, Subasinghe C, Hélène C, Cohen JS, Toulmé JJ

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.

MeSH Terms
Animals Base Sequence Binding, Competitive Cell-Free System Globins/genetics,metabolism Molecular Sequence Data Nucleic Acid Hybridization Oligonucleotide Probes/chemical synthesis,pharmacology Oocytes/metabolism Protein Biosynthesis RNA RNA, Antisense RNA, Messenger/genetics,metabolism Rabbits Reticulocytes/metabolism Ribonucleases/metabolism Thermodynamics Triticum/genetics Xenopus/genetics
Chemicals
Oligonucleotide Probes RNA, Antisense RNA, Messenger RNA Globins Ribonucleases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Cazenave C
Laboratoire de Biophysique, INSERM U201, Paris, France.
Stein C A
Loreau N
Thuong N T
Neckers L M
Subasinghe C
Hélène C
Cohen J S
Toulmé J J
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.11.4255
Published
1989-06-12
Pages
4255-73
Language
English
Region
England
NLM ID
0411011
PMCID
PMC317933
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/17.11.4255 PMC Full Text View journal details
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