Incorporation of multiple sequential pseudothymidines by DNA polymerases and their impact on DNA duplex structure.
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.
Havemann
Stephanie A
SA
Department of Microbiology & Cell Science, Space Life Sciences Laboratory, Kennedy Space Center, University of Florida, FL, USA.
Hoshika
Shuichi
S
Hutter
Daniel
D
Benner
Steven A
SA
D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D006027:Glycosides; D009005:Monosaccharides / Q000737:chemistry* / Q000378:metabolism; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D011741:Pyrimidine Nucleosides / Q000737:chemistry* / Q000378:metabolism; D019914:Taq Polymerase / Q000378:metabolism
C438934:C-glycoside; D006027:Glycosides; D009005:Monosaccharides; D009841:Oligonucleotides; D011741:Pyrimidine Nucleosides; C096786:pseudothymidine; D004247:DNA; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase