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J. Biol. Chem., Vol. 275, Issue 51, 40601-40604, December 22, 2000

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Singlet Oxygen Induces Oxidation of Cellular DNA^*

Jean-Luc Ravanat, Paolo Di Mascio, Glaucia R. Martinez, Marisa H. G. Medeiros, and Jean Cadet^§

From the Laboratoire "Lésions des Acides Nucléiques" Service de Chimie Inorganique et Biologique, UMR CNRS 5046, Département de Recherche Fondamentale sur la Matière Condensée, CEA Grenoble, 17 Avenue des Martyrs, F-38054 Grenoble Cedex 9, France and the  Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, CP 26077, CEP 05513-970, São Paulo, SP, Brazil

The aim of the present work was to evaluate the potential for ¹O₂ to induce oxidation of cellular DNA. For this purpose cells were incubated in the presence of a water-soluble endoperoxide whose thermal decomposition leads to the formation of singlet oxygen. Thereafter, DNA was extracted and the level of several modified DNA bases was determined by HPLC analysis coupled to a tandem mass spectrometric detection. A significant increase in the level of 8-oxo-7,8-dihydro-2'-deoxyguanosine was observed upon incubation of the cells with the chemical generator of ¹O₂, whereas the level of the other DNA bases measured remained unchanged. To demonstrate that singlet oxygen is directly involved in the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine, the corresponding ¹⁸O-labeled endoperoxide was used. Incubation of the cells with such a generator of ¹⁸O-labeled singlet oxygen results in the formation of ¹⁸O-labeled 8-oxo-7,8-dihydro-2'-deoxyguanosine in the nuclear DNA. This result clearly demonstrates that singlet oxygen, when released within cells, is able to directly oxidize cellular DNA.

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