Enzymatic incorporation in DNA of 1,5-anhydrohexitol nucleotides.

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.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Vastmans K,Pochet S,Peys A,Kerremans L,Van Aerschot A,Hendrix C,Marlière P,Herdewijn P

doi

10.1021/bi001297g

subject

Has Abstract

pub_date

2000-10-24 00:00:00

pages

12757-65

issue

42

eissn

0006-2960

issn

1520-4995

pii

bi001297g

journal_volume

39

pub_type

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