Intrastrand G-U cross-links generated by the oxidation of guanine in 5′-d(GCU) and 5′-r(GCU)

Conor Crean, Nicholas Geacintov, Vladimir Shafirovich

Research output: Contribution to journalArticle

Abstract

It has been suggested that carbonate radical anions are biologically important because they may be produced during the inflammatory response. The carbonate radicals can selectively oxidize guanine in DNA and RNA by one-electron transfer mechanisms and the guanine radicals thus formed decay by diverse competing pathways with other free radicals or nucleophiles. Using a photochemical method to generate CO3{radical dot}- radicals in vitro, we compare the distributions of products initiated by the one-electron oxidation of guanine in the trinucleotides 5′-r(GpCpU) and 5′-d(GpCpU) in aqueous buffer solutions (pH 7.5). Similar distributions of stable end products identified by LC-MS/MS methods were found in both cases. The guanine oxidation products include the diastereomeric pair of spiroiminodihydantoin (Sp) and 2,5-diamino-4H-imidazolone (Iz). In addition, intrastrand cross-linked products involving covalent bonds between the G and the U bases (G*CU*) were also found, although with different relative yields in the 2′-deoxy- and the ribotrinucleotides. The positive-ion MS/MS spectra of the 5′-r(G*pCpU*) and 5′-d(G*pCpU*) products clearly indicate the presence of covalently linked G*-U* products that have a mass smaller by 2 Da than the sum of the G and U bases in both types of trinucleotides. The 5′-d(G*CU*) cross-linked product was further characterized by 1D and 2D NMR methods that confirm its cyclic structure in which the guanine C8 atom is covalently linked to the uracil N3 atom.

Original languageEnglish (US)
Pages (from-to)1125-1134
Number of pages10
JournalFree Radical Biology and Medicine
Volume45
Issue number8
DOIs
StatePublished - Oct 15 2008

Fingerprint

Guanine
Oxidation
Carbonates
Electrons
Atoms
Nucleophiles
Covalent bonds
Uracil
Free Radicals
Anions
Buffers
Positive ions
Nuclear magnetic resonance
RNA
Ions
DNA

Keywords

  • Carbonate radical anion
  • Cross-linked DNA
  • DNA
  • Free radicals
  • Guanine
  • Oxidation
  • RNA
  • Uracil

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

Cite this

Intrastrand G-U cross-links generated by the oxidation of guanine in 5′-d(GCU) and 5′-r(GCU). / Crean, Conor; Geacintov, Nicholas; Shafirovich, Vladimir.

In: Free Radical Biology and Medicine, Vol. 45, No. 8, 15.10.2008, p. 1125-1134.

Research output: Contribution to journalArticle

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