Structural origins of bulky oxidative DNA adducts (type II I-compounds) as deduced by oxidation of oligonucleotides of known sequence.
Bulky DNA adducts, previously termed type II I-compounds, are detected by 32P-postlabeling following treatment of DNA with several Fenton-type oxygen radical-generating reagents, i.e., mixtures of Fe(II) or Ni(II) and H2O2. In an attempt to characterize the chemical nature and mechanism(s) of formation of these novel adducts, 16 single-stranded deoxyribooligonucleotides (20- and 21-mers) of known sequence were oxidized with Fe(II) or Ni(II) and H2O2, and the products were analyzed by 32P-postlabeling. Eight adducts were obtained reproducibly by oxidation of DNA and test oligonucleotides in a sequence-dependent manner. One major adduct (2) was formed only if the test oligonucleotide contained two adjacent adenine residues. Similarly, adducts 3 and 8 specifically originated in AC and CA sequences, respectively. Adduct 6 required a 5'-C-purine-3' sequence. On the other hand, GN sequences (where N is any normal nucleotide) gave rise to adduct 1, another major product, and adduct 7. Similarly, adducts 4 and 5 were produced by the oxidation of AN sequences. These observations are most readily explained if the oxidation reactions caused intrastrand cross-links between adjacent nucleotides, leading to dimer formation. The observation that adducts 1, 4, 5, and 7 did not require a specific 3'-nucleotide was consistent with the notion that these nucleotides lacked a 3'-base, suggesting the presence of a 5'-->3' purine-sugar cross-linked in the oxidized products. The majority of the lesions came from AA and 5'-purine-N-3' sequences. The effects of Fe(II) and Ni(II) were qualitatively similar; however, higher yields of products were observed with Fe(II) as the catalyst. The definition of the chemical origins of these bulky DNA modifications, which represent a new type of DNA damage, is expected to contribute to a better understanding of the mechanism of metal carcinogenesis and to shed light upon the origins of certain endogenous DNA lesions. Recently, some of the major oxidative DNA adducts characterized here were detected by 32P-postlabeling in the renal DNA of male rats treated with ferric nitrilotriacetate, a known potent prooxidative kidney carcinogen in these animals.
Randerath
K
K
Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77030, USA.
Randerath
E
E
Smith
C V
CV
Chang
J
J
D001345:Autoradiography; D001482:Base Composition; D001483:Base Sequence; D002384:Catalysis; D018736:DNA Adducts / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D009532:Nickel / Q000494:pharmacology; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D010761:Phosphorus Radioisotopes; D013329:Structure-Activity Relationship
D018736:DNA Adducts; D009841:Oligonucleotides; D010761:Phosphorus Radioisotopes; D009532:Nickel