Oligonucleotide-directed switching of DNA polymerases to a dead-end track

Biochemistry. 1995 Jul 18;34(28):9193-9. doi: 10.1021/bi00028a032.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA / genetics
  • DNA / metabolism
  • DNA Replication / drug effects*
  • DNA-Directed DNA Polymerase / metabolism*
  • Humans
  • Molecular Sequence Data
  • Oligonucleotides / pharmacology*
  • Plasmids / genetics
  • Polymerase Chain Reaction
  • Receptors, Interleukin-2 / genetics
  • Taq Polymerase

Substances

  • Oligonucleotides
  • Receptors, Interleukin-2
  • DNA
  • Taq Polymerase
  • bacteriophage T7 induced DNA polymerase
  • DNA-Directed DNA Polymerase