Investigation of the DNA-dependent cyclohexenyl nucleic acid polymerization and the cyclohexenyl nucleic acid-dependent DNA polymerization.
DNA polymerases from different evolutionary families [Vent (exo-) DNA polymerase from the B-family polymerases, Taq DNA polymerase from the A-family polymerases and HIV reverse transcriptase from the reverse transcriptase family] were examined for their ability to incorporate the sugar-modified cyclohexenyl nucleoside triphosphates. All enzymes were able to use the cyclohexenyl nucleotides as a substrate. Using Vent (exo-) DNA polymerase and HIV reverse transcriptase, we were even able to incorporate seven consecutive cyclohexenyl nucleotides. Using a cyclohexenyl nucleic acid (CeNA) template, all enzymes tested were also able to synthesize a short DNA fragment. Since the DNA-dependent CeNA polymerization and the CeNA-dependent DNA polymerization is possible to a limited extend, we suggest CeNA as an ideal candidate to use in directed evolution methods for the development of a polymerase capable of replicating CeNA.
Kempeneers
Veerle
V
Laboratory for Medicinal Chemistry, Rega Institute for Medical Research Minderbroedersstraat 10, B-3000 Leuven, Belgium.
Renders
Marleen
M
Froeyen
Matheus
M
Herdewijn
Piet
P
D001704:Biopolymers / Q000096:biosynthesis / Q000737:chemistry; D003510:Cyclohexanes / Q000737:chemistry*; D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D019020:Directed Molecular Evolution; D007700:Kinetics; D009711:Nucleotides / Q000737:chemistry* / Q000378:metabolism*; D013698:Templates, Genetic
D001704:Biopolymers; D003510:Cyclohexanes; D009711:Nucleotides; D004247:DNA; D004259:DNA-Directed DNA Polymerase