Hybridization specificity, enzymatic activity and biological (Ha-ras) activity of oligonucleotides containing 2,4-dideoxy-beta-D-erythro-hexopyranosyl nucleosides.
Antisense oligonucleotides with a 2,4-dideoxyhexopyranosyl nucleoside incorporated at the 3'-end and at a mutation site of the Ha-ras oncogene mRNA were synthesized. Melting temperature studies revealed that an A*-G mismatch is more stable than an A*-T mismatch with these hexopyranosyl nucleosides incorporated at the mutation site. The oligonucleotides are stable against enzymatic degradation. RNase H mediated cleavage studies revealed selective cleavage of mutated Ha-ras mRNA. The oligonucleotide containing two pyranose nucleosides at the penultimate position activates RNase H more strongly than natural oligonucleotides. No correlation, however, was found between DNA - DNA or RNA - DNA melting temperatures and RNase H mediated cleavage capacity. Although the A*-G mismatch gives more stable hybridization than the A*-T base pairing, only the oligonucleotides containing an A*-T base pair are recognized by RNase H. This modification is situated 3 base pairs upstream to the cleavage site. Finally, the double pyranose modified oligonucleotide was able to reduce the growth of T24 cells (bladder carcinoma) while the unmodified antisense oligonucleotide was not.
Augustyns
K
K
Rega Institute for Medical Research, Laboratory of Medicinal Chemistry, Katholieke Universiteit Leuven, Belgium.
Godard
G
G
Hendrix
C
C
Van Aerschot
A
A
Rozenski
J
J
Saison-Behmoaras
T
T
Herdewijn
P
P
D001482:Base Composition; D001483:Base Sequence; D003837:Deoxy Sugars / Q000737:chemistry*; D015224:Dideoxynucleosides / Q000138:chemical synthesis / Q000737:chemistry*; D011905:Genes, ras; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009693:Nucleic Acid Hybridization; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000737:chemistry*; D012313:RNA; D016914:Ribonuclease H / Q000378:metabolism; D013696:Temperature; D014407:Tumor Cells, Cultured
D003837:Deoxy Sugars; D015224:Dideoxynucleosides; D016376:Oligonucleotides, Antisense; D012313:RNA; C041005:2,4-dideoxyhexopyranose; D016914:Ribonuclease H