Independent generation and characterization of a C2'-oxidized abasic site in chemically synthesized oligonucleotides

J Org Chem. 2004 Sep 3;69(18):6100-4. doi: 10.1021/jo049033d.

Abstract

Abasic lesions, which are formed endogenously and as a consequence of exogenous agents, are lethal and mutagenic. Hydrogen atom abstraction from C2' in DNA under aerobic conditions produces an oxidized abasic lesion (C2-AP), along with other forms of DNA damage. The effects of C2-AP on DNA structure and function are not well understood. A method for the solid-phase synthesis of oligonucleotides containing C2-AP lesions is reported. The lesion is released via periodate oxidation of a triol containing a vicinal diol. The triol is introduced via a phosphoramidite that is compatible with standard oligonucleotide synthesis and deprotection conditions. UV-melting studies indicate that the C2-AP lesion has a comparable effect on the thermal stability of duplex DNA as other abasic lesions. The C2-AP lesion is rapidly cleaved by piperidine at 90 degrees C. However, cleavage by NaOH (0.1 M, 37 degrees C) shows that C2-AP is considerably less labile (t(1/2) = 3.3 +/- 0.2 h) than other abasic lesions.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • DNA / chemistry*
  • DNA Damage*
  • Models, Molecular
  • Molecular Structure
  • Oligonucleotides / chemical synthesis*
  • Oxidation-Reduction
  • Oxidative Stress*
  • Piperidines / chemistry
  • Time Factors

Substances

  • Oligonucleotides
  • Piperidines
  • DNA