Crosslinking reactions of 4-amino-6-oxo-2-vinylpyrimidine with guanine derivatives and structural analysis of the adducts

Nucleic Acids Res. 2015 Sep 18;43(16):7717-30. doi: 10.1093/nar/gkv797. Epub 2015 Aug 5.

Abstract

DNA interstrand crosslinks (ICLs) are the primary mechanism for the cytotoxic activity of many clinical anticancer drugs, and numerous strategies for forming ICLs have been developed. One such method is using crosslink-forming oligonucleotides (CFOs). In this study, we designed a 4-amino-6-oxo-2-vinylpyrimidine (AOVP) derivative with an acyclic spacer to react selectively with guanine. The AOVP CFO exhibited selective crosslinking reactivity with guanine and thymine in DNA, and with guanine in RNA. These crosslinking reactions with guanine were accelerated in the presence of CoCl2, NiCl2, ZnCl2 and MnCl2. In addition, we demonstrated that the AOVP CFO was reactive toward 8-oxoguanine opposite AOVP in the duplex DNA. The structural analysis of each guanine and 8-oxoguanine adduct in the duplex DNA was investigated by high-resolution NMR. The results suggested that AOVP reacts at the N2 amine in guanine and at the N1 or N2 amines in 8-oxoguanine in the duplex DNA. This study demonstrated the first direct determination of the adduct structure in duplex DNA without enzyme digestion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cross-Linking Reagents / chemistry*
  • DNA / chemistry
  • DNA Adducts / chemistry*
  • Guanine / analogs & derivatives
  • Guanine / chemistry*
  • Metals / chemistry
  • Oligonucleotides / chemical synthesis
  • Pyrimidinones / chemistry*
  • RNA / chemistry
  • Thymine / chemistry
  • Vinyl Compounds / chemistry*

Substances

  • Cross-Linking Reagents
  • DNA Adducts
  • Metals
  • Oligonucleotides
  • Pyrimidinones
  • Vinyl Compounds
  • Guanine
  • RNA
  • DNA
  • Thymine