Oligonucleotide-directed switching of DNA polymerases to a dead-end track.
During DNA replication, the presence of oligonucleotides with partial homology to the template strand was shown to induce a switch of the polymerase from the normal template to the oligonucleotide. The latter acted as a dead-end template and led to abortive replication. The only prerequisite was that the oligonucleotide could form 7-9 base pairs with the newly synthesized DNA strand in order to switch templates. The switch occurred when base pairing of the oligonucleotide could take place with the 3'-end of the newly synthesized strand. These results show that oligonucleotides used in antisense or antigene strategies could have unexpected effects on replication. In addition, oligonucleotide-directed abortive replication might play an inhibitory role during PCR experiments on long DNA templates and lead to the amplification of truncated fragments.
Guieysse
A L
AL
Laboratoire de Biophysique, INSERM U.201--CNRS URA 481, Muséum National d'Histoire Naturelle, Paris, France.
Praseuth
D
D
Grigoriev
M
M
Harel-Bellan
A
A
Hélène
C
C
D001483:Base Sequence; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000494:pharmacology*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D015375:Receptors, Interleukin-2 / Q000235:genetics; D019914:Taq Polymerase
D009841:Oligonucleotides; D015375:Receptors, Interleukin-2; D004247:DNA; D019914:Taq Polymerase; C034970:bacteriophage T7 induced DNA polymerase; D004259:DNA-Directed DNA Polymerase