Incorporation of multiple sequential pseudothymidines by DNA polymerases and their impact on DNA duplex structure

Nucleosides Nucleotides Nucleic Acids. 2008 Mar;27(3):261-78. doi: 10.1080/15257770701853679.

Abstract

Thermal denaturation and circular dichroism studies suggested that multiple (up to 12), sequential pseudothymidines, a representative C-glycoside, do not perturb the structure of a representative DNA duplex. Further, various Family A and B DNA polymerases were found to extend a primer by incorporating four sequential pseudothymidine triphosphates, and then continue the extension to generate full-length product. Detailed studies showed that Taq polymerase incorporated up to five sequential C-glycosides, but not more. These results constrain architectures for sequencing, quantitating, and analyzing DNA analogs that exploit C-glycosides, and define better the challenge of creating a synthetic biology using these with natural polymerases.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Circular Dichroism
  • DNA / chemistry*
  • DNA / metabolism
  • DNA-Directed DNA Polymerase / metabolism*
  • Glycosides
  • Monosaccharides / chemistry*
  • Monosaccharides / metabolism
  • Nucleic Acid Conformation
  • Nucleic Acid Denaturation
  • Oligonucleotides / chemical synthesis
  • Oligonucleotides / chemistry*
  • Oligonucleotides / metabolism
  • Pyrimidine Nucleosides / chemistry*
  • Pyrimidine Nucleosides / metabolism
  • Taq Polymerase / metabolism

Substances

  • C-glycoside
  • Glycosides
  • Monosaccharides
  • Oligonucleotides
  • Pyrimidine Nucleosides
  • pseudothymidine
  • DNA
  • Taq Polymerase
  • DNA-Directed DNA Polymerase