Incorporation of 2'-deoxy-2'-isonucleoside 5'-triphosphates (iNTPs) into DNA by A- and B-family DNA polymerases with different recognition mechanisms.
Recently, α-L-threofuranosyl nucleoside 3'-triphosphates (tNTPs) have been reported to be incorporated into DNA by DNA polymerases. Isonucleosides especially the 2'-deoxy-2'-isonucleosides, would be considered regioisomers of α-L-threofuranosyl nucleosides. Therefore, we investigated the synthesis of 2'-deoxy-2'-isonucleoside 5'-triphosphates (iNTPs) having the four natural nucleobases and their incorporation into primer-template duplexes consisting of oligonucleotides containing natural 2'-deoxyribonucleosides and 2'-deoxy-2'-isonucleosides by using primer-extension reactions. We found that Klenow fragment (exo-; an A-family DNA polymerase) has strict recognition of the shape of nucleoside 5'-triphosphates and Therminator (a B-family DNA polymerase) has strict recognition of the shape of primer-template complexes, especially two base pairs upstream of the primer 3' terminus.
Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
Takaaki
T
Ogino
Kousuke
K
Sato
Akira
A
Matsuda
D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D004256:DNA Polymerase I / Q000378:metabolism; D017931:DNA Primers / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D003854:Deoxyribonucleotides / Q000378:metabolism; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*
D017931:DNA Primers; D003854:Deoxyribonucleotides; D009841:Oligonucleotides; D004247:DNA; D004256:DNA Polymerase I; D004259:DNA-Directed DNA Polymerase