Antisense Oligonucleotides Containing Modified Bases Inhibit in Vitro Translation of Leishmania amazonensis mRNAs by Invading the Mini-exon Hairpin

Complementary oligodeoxynucleotides (ODNs) that contain 2-aminoadenine and 2-thiothymine interact weakly with each other but form stable hybrids with unmodified complements. These selectively binding complementary (SBC) agents can invade duplex DNA and hybridize to each strand (Kutyavin, I. V., Rhin...

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Bibliographic Details
Published in:The Journal of biological chemistry 1999-03, Vol.274 (12), p.8191-8198
Main Authors: Compagno, D, Lampe, J N, Bourget, C, Kutyavin, I V, Yurchenko, L, Lukhtanov, E A, Gorn, V V, Gamper, Jr, H B, Toulmé, J J
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Chromosome Mapping
Escherichia coli
Exons
Hot Temperature
Leishmania - genetics
Leishmania amazonensis
Molecular Sequence Data
Nucleic Acid Conformation
Oligonucleotides, Antisense - metabolism
Protein Biosynthesis
RNA, Messenger - metabolism
RNA, Protozoan - metabolism
Thionucleotides - metabolism
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Summary:Complementary oligodeoxynucleotides (ODNs) that contain 2-aminoadenine and 2-thiothymine interact weakly with each other but form stable hybrids with unmodified complements. These selectively binding complementary (SBC) agents can invade duplex DNA and hybridize to each strand (Kutyavin, I. V., Rhinehart, R. L., Lukhtanov, E. A., Gorn, V. V., Meyer, R. B., and Gamper, H. B. (1996) Biochemistry 35, 11170–11176). Antisense ODNs with similar properties should be less encumbered by RNA secondary structure. Here we show that SBC ODNs strand invade a hairpin in the mini-exon RNA of Leishmania amazonensis and that the resulting heteroduplexes are substrates for Escherichia coli RNase H. SBC ODNs either with phosphodiester or phosphorothioate backbones form more stable hybrids with RNA than normal base (NB) ODNs. Optimal binding was observed when the entire hairpin sequence was targeted. Translation of L. amazonensis mRNA in a cell-free extract was more efficiently inhibited by SBC ODNs complementary to the mini-exon hairpin than by the corresponding NB ODNs. Nonspecific protein binding in the cell-free extract by phosphorothioate SBC ODNs rendered them ineffective as antisense agents in vitro . SBC phosphorothioate ODNs displayed a modest but significant improvement of leishmanicidal properties compared with NB phosphorothioate ODNs.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.274.12.8191