Site‐specific mutagenicity of stereochemically defined 1,N²‐deoxyguanosine adducts of trans‐4‐hydroxynonenal in mammalian cells

Abstract

Trans-4-hydroxynonenal (HNE) is a toxic compound produced endogenously during lipid peroxidation. HNE is a potent electrophile that is reactive with both proteins and nucleic acids. HNE preferentially reacts with deoxyguanosine to form four stereoisomeric HNE-deoxyguanosine (HNE-dG) adducts: (6R, 8S, 11R), (6S, 8R, 11S), (6R, 8S, 11S), and (6S, 8R, 11R). These adducts were synthesized into 12-mer oligodeoxynucleotides, inserted into a DNA shuttle vector and evaluated for the ability of each stereoisomer to induce mutagenesis when replicated through mammalian cells. The resultant mutagenicity of these adducts was related to their stereochemistry, in that two of the HNE-dG adducts, (6R, 8S, 11R) and (6S, 8R, 11S), were significantly more mutagenic than the (6R, 8S, 11S) and (6S, 8R, 11R) HNE-dG adducts. These data conclusively demonstrate that HNE-derived DNA adducts can be mutagenic in mammalian cells and their ability to cause mutations is dictated by their stereochemistry.

Replication past a trans-4-Hydroxynonenal Minor-Groove Adduct by the Sequential Action of Human DNA Polymerases ι and κ

“…The observation of a more proficient extension by Pol from a C opposite the 11R isomer than from a C opposite the 11S isomer may be because of the greater prevalence of the ring-open configuration in the 11R isomer than the 11S isomer, and that could also account for Pol's ability to extend from a C opposite the 11R isomer. The assumption that ring opening is somewhat more efficient for the 11R isomer is in a good agreement with two observations from other studies: (i) melting experiments showed that a duplex oligodeoxynucleotide containing the 11S HNE-dG is significantly more destabilized than the 11R-containing duplex (25) and (ii) whereas the 11R isomer was essentially nonmutagenic in COS-7 cells, the 11S adduct yielded approximately 4% mutations (6).…”

Trans -4-hydroxy-2-nonenal inhibits nucleotide excision repair in human cells: A possible mechanism for lipid peroxidation-induced carcinogenesis

“…Previously, we and other (13,14) have found that the 4-HNE-dG adduct induces mainly G:C to T:A transversions in human cells, and 4-HNE forms DNA adducts preferentially at codon 249 of p53 gene (15), a mutational hotspot in hepatocellular carcinoma and smoke-related lung cancer (16). Recently, it has been shown that 4-HNE can indeed induce a high frequency of G:C to T:A mutations at codon 249 of the p53 gene (37).…”

“…It has been found that 4-HNE can interact with DNA to form 4-HNE-dG adduct, a bulky exocyclic DNA adduct, which has been found in various normal tissues of humans and rats (1,9,12). We and others (13,14) have recently found that 4-HNE-dG adduct is a strong mutagen and induces mainly G:C to T:A mutations in human cells. We have also found that 4-HNE-dG adduct preferentially form at -GAGGC͞A-sequences in the p53 gene, including codon 249 (15), a mutational hotspot in human hepatocellular carcinoma and cigarette smokerelated lung cancer (16).…”

Damage of DNA and proteins by major lipid peroxidation products in genome stability

“…Etheno-type adduct of HNE to guanosine, 1,N 2 ε-guanosine, and propano-type cyclic 1,N 2 -propanodeoxyguanine were shown to occur as four diastereoisomers of tetracyclic structure (purine condensed with ethano and furano rings) [40,41]. These isomers were shown to have different mutagenic potency [42]. Other cyclic DNA adducts deriving from acrolein and crotonaldehyde occur in stereomeric forms as well [43,44].…”