Enzymatic Incorporation in DNA of 1,5-Anhydrohexitol Nucleotides
- 28 September 2000
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 39 (42) , 12757-12765
- https://doi.org/10.1021/bi001297g
Abstract
The ability of several DNA polymerases to catalyze the template-directed synthesis of duplex oligonucleotides containing a base pair between a nucleotide with anhydrohexitol ring and its natural complement has been investigated. All DNA polymerases were able to accept the chemically synthesized anhydrohexitol triphosphate as substrate and to catalyze the incorporation of one anhydrohexitol nucleotide. However, only family B DNA polymerases succeeded in elongating the primer after the incorporation of an anhydrohexitol nucleotide. In this family, Vent (exo-) DNA polymerase is the most successful one and was therefore selected for further investigation. Results revealed that at high enzyme concentrations six hATPs could be incorporated; however, a selective incorporation proved only feasible under experimental conditions where no more than two analogues could be inserted. Also the synthesis of a mixed HNA−DNA sequence was examined. Kinetic parameters for incorporation of one anhydrohexitol adenine nucleoside were similar to those of its natural analogue.Keywords
This publication has 12 references indexed in Scilit:
- Cloning and Characterization of a Family B DNA Polymerase from the Hyperthermophilic Crenarchaeon Pyrobaculum islandicumJournal of Bacteriology, 2000
- 2-Aza-2′-deoxyadenosine: Synthesis, Base-Pairing Selectivity, and Stacking Properties of OligonucleotidesChemistry – A European Journal, 2000
- Efforts toward Expansion of the Genetic Alphabet: DNA Polymerase Recognition of a Highly Stable, Self-Pairing Hydrophobic BaseJournal of the American Chemical Society, 1999
- How E. coli DNA polymerase I (klenow fragment) distinguishes between deoxy- and dideoxynucleotidesJournal of Molecular Biology, 1998
- Acyclic nucleoside triphosphate analogs as terminators in biocatalytic DNA replicationBioorganic & Medicinal Chemistry Letters, 1997
- Crystal structure of a thermostable Bacillus DNA polymerase l large fragment at 2.1 Å resolutionStructure, 1997
- An Approach to Random Mutagenesis of DNA Using Mixtures of Triphosphate Derivatives of Nucleoside AnaloguesJournal of Molecular Biology, 1996
- Synthesis, Biological Evaluation, and Structure Analysis of a Series of New 1,5-Anhydrohexitol NucleosidesJournal of Medicinal Chemistry, 1995
- FIDELITY MECHANISMS IN DNA REPLICATIONAnnual Review of Biochemistry, 1991
- Direct sequencing of polymerase chain reaction amplified DNA fragments through the incorporation of deoxynucleoside α-thiotriphosphatesNucleic Acids Research, 1988