Incorporation by chemical synthesis and characterization of deoxyribosylformylamine into DNA.
2-deoxyribosylformylamine is a major oxidative DNA damage type which occurs upon the action of ionizing radiation on DNA. The protected 2-deoxyribosylformylamine phosphoramidite was synthesized and used in conjunction with previously reported alkali labile base protected phosphoramidites ('PAC phosphoramidites') for the preparation of oligodeoxyribonucleotides containing this lesion. Final deprotection of the oligonucleotides was performed under mild alkaline conditions to preserve the integrity of the fragile defect. The presence of formylamino deoxyribosyl residue was confirmed by FAB mass spectrometry sequencing. Oligonucleotides bearing deoxyribosyl formylamine were used as templates for studying in vitro replication. They direct the insertion of guanine or induce a deletion opposite the lesion.
Guy
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Département de Recherche Fondamentale de la Matière Condensée, Centre d'Etudes Nucléaires de Grenoble, France.
Duplaa
A M
AM
Ulrich
J
J
Téoule
R
R
D001483:Base Sequence; D004249:DNA Damage; D004261:DNA Replication / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003855:Deoxyribose / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D006147:Guanine / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis* / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000138:chemical synthesis* / Q000378:metabolism; D019914:Taq Polymerase
D009838:Oligodeoxyribonucleotides; D009943:Organophosphorus Compounds; D003855:Deoxyribose; C071599:2-deoxyribosylformylamine; D006147:Guanine; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase