Repair and replication of oxidized DNA bases using modified oligodeoxyribonucleotides.
Modified oligodeoxyribonucleotides (ODNs) are powerful tools to assess the biological significance of oxidized lesions to DNA. For this purpose, we developed original synthetical pathways for the site-specific insertion of several oxidized bases into DNA fragments. Thus, the chemical solid-phase synthesis of ODNs using original strategies of protection and mild conditions of deprotection, as well as a specific post-oxidation approach of an unique nucleoside residue within the sequence have been applied. These two approaches of preparation allowed us to have access to a set of modified ODNs that contain a single modified nucleoside, i.e., N-(2-deoxy-beta-D-erythro-pentofuranosyl)formylamine (dF), 5-hydroxy-2'-deoxycytidine (5-OHdCyd), thymidine glycol (dTg), 5,6-dihydrothymidine (DHdThd), 2,2-diamino-4-[(2-deoxy-beta-D-erythro-pentofuranosyl)-amino]-5(2H)- oxazolone (dZ), N-(2-deoxy-beta-D-erythro-pentofuranosyl)cyanuric acid (dY), 5',8-cyclo-2'-deoxyguanosine (cyclodGuo) and 5',8-cyclo-2'-deoxyadenosine (cyclodAdo). The substrates were used to investigate recognition and removal of the lesions by bacterial DNA N-glycosylases, including endonuclease III (endo III) and Fapy glycosylase (Fpg). In addition, the DNA polymerase-mediated nucleotide incorporation opposite the damage was determined using modified ODNs as templates.
Gasparutto
D
D
Laboratoire Lésions des Acides Nucléiques, C.E.A-Grenoble, France.
Bourdat
A G
AG
D'Ham
C
C
Duarte
V
V
Romieu
A
A
Cadet
J
J
D020029:Base Pairing; D004247:DNA / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D045647:DNA Glycosylases; D004260:DNA Repair; D004261:DNA Replication / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000235:genetics; D010084:Oxidation-Reduction; D013379:Substrate Specificity
D009838:Oligodeoxyribonucleotides; D004247:DNA; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases