A phosphorothioate oligonucleotide blocks reverse transcription via an antisense mechanism.
We have studied the inhibition by a phosphorothioate oligodeoxynucleotide (17PScap) of cDNA synthesis performed by either avian or murine reverse transcriptase. Three different mechanisms of inhibition were identified: at low concentrations (< 100 nM), the cleavage of the RNA template by the retroviral RNase H at the level of the RNA/17PScap duplex accounted for most of the effect, whereas hybrid-arrested cDNA synthesis by an RNase H-independent mechanism marginally contributed to the inhibition. Both mechanisms were sequence-specific. Above 100 nM, the overall cDNA synthesis was reduced in a non-specific manner.
Boiziau
C
C
Laboratoire de Biophysique Moléculaire, INSERM U 386, Université de Bordeaux II, France.
Moreau
S
S
Toulmé
J J
JJ
D000818:Animals; D001483:Base Sequence; D001717:Birds; D018076:DNA, Complementary / Q000096:biosynthesis; D006868:Hydrolysis; D051379:Mice; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism; D013698:Templates, Genetic; D013873:Thionucleotides / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000235:genetics*
D018076:DNA, Complementary; D016376:Oligonucleotides, Antisense; D013873:Thionucleotides; D012194:RNA-Directed DNA Polymerase; D016914:Ribonuclease H