In vitro reaction of ethylene oxide with DNA and characterization of DNA adducts

Li, Fujun; Segal, Alvin; Solomon, Jerome J.

doi:10.1016/0009-2797(92)90090-8

Cited by 77 publications

(77 citation statements)

References 44 publications

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“…A fraction was collected from 17 to 20 min (residual NNs had the following retention times: 2'-deoxycytidine-3'-monophosphate, 4.3 min; 2'-deoxyguanosine-3'-monophosphate, 5.0 min; 2'-deoxythymidine-3'-monophosphate, 6.6 min; 2'-deoxyadenosine-3'-monophosphate, 7.7 min). Enriched adducts were postlabeled to 5' position by T4 kinase; the formed bisphosphates were treated with nuclease P1 resulting in 5' monophosphate adducts.…”

Section: Methodsmentioning

confidence: 99%

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Inhalation Exposure of Rats and Mice to 1,3-Butadiene Induces N 6 -Adenine Adducts of Epoxybutene Detected by 32 P-Postlabeling and HPLC

Koivisto¹,

Adler²,

Sorsa³

et al. 1996

Environmental Health Perspectives

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In this paper we report DNA binding of butadiene monoepoxide, a first metabolite of 1,3-butadiene catalyzed by monooxygenases. We prepared alkylated purines as marker compounds for 32P-postlabeling and electrochemical analysis and developed methods to measure the corresponding products. The traditional postlabeling assay was modified by incorporating a solid phase extraction column and high-performance liquid chromatography (HPLC) enrichment steps to the assay prior to labeling. The final analysis of adducted N6 adenines is based on two dimensional thin-layer chromatography (TLC) and an on-line HPLC/radioactivity analysis. The qualitative and quantitative results are based on positively identified marker compounds. Alkylated N7 guanines were released from DNA by neutral thermal hydrolysis, prepurified by HPLC, and analyzed by HPLC with a sensitive electrochemical detection procedure. By using these methods, we found alkylation of calf thymus DNA exposed to butadiene monoepoxide in vitro at adenine N6 and guanine N7 sites. Analysis of lung DNA samples from mice and rats exposed to butadiene through inhalation showed that adenine N6 adducts were formed in vivo in a dose responsive manner. Environ Health Perspect 1 04(Suppl 3): 655-657 (1996)

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Section: Methodsmentioning

confidence: 99%

“…A recent epidemiological study in the BD industry indicates an elevated mortality of lymphatic and reticular sarcoma among the production workers (3). Small alkylating agents react mainly with the N7 position of guanine, but other alkylation sites in DNA are also detected in vitro (4,5). Goodrow et.…”

Section: Introductionmentioning

confidence: 99%

Inhalation Exposure of Rats and Mice to 1,3-Butadiene Induces N 6 -Adenine Adducts of Epoxybutene Detected by 32 P-Postlabeling and HPLC

Koivisto¹,

Adler²,

Sorsa³

et al. 1996

Environmental Health Perspectives

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“…The mutagenicity and carcinogenicity of EO is attributed to reaction with DNA, leading to the formation of multiple 2-hydroxyethyl adducts (11,12). The most abundant product, N7-(2-hydroxyethyl)guanine (N7-HEG), readily depurinates, leaving abasic sites with miscoding potential (13,14).…”

Section: Introductionmentioning

confidence: 99%

Dose-Response Relationships for N7-(2-Hydroxyethyl)Guanine Induced by Low-Dose [14C]Ethylene Oxide: Evidence for a Novel Mechanism of Endogenous Adduct Formation

et al. 2009

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Ethylene oxide (EO) is widely used in the chemical industry and is also formed in humans through the metabolic oxidation of ethylene, generated during physiologic processes. EO is classified as a human carcinogen and is a direct acting alkylating agent, primarily forming N7-(2-hydroxyethyl)guanine (N7-HEG). To conduct accurate human risk assessments, it is vital to ascertain the relative contribution of endogenously versus exogenously derived DNA damage and identify the sources of background lesions. We have therefore defined in vivo dose-response relationships over a concentration range relevant to human EO exposures using a dual-isotope approach. By combining liquid chromatography-tandem mass spectrometry and high-performance liquid chromatographyaccelerator mass spectrometry analysis, both the endogenous and exogenous N7-HEG adducts were quantified in tissues of [ 14 C]EO-treated rats.

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“…3-HA-dU is formed after initial alkylation at the N3 position of dC, followed by rapid hydrolytic deamination to 3-HA-dU (34,37,38). The hydroxyalkyl group of 3-HA-dU occupies a central Watson-Crick hydrogen-bonding position and is likely to disrupt normal base pairing.…”

Section: Introductionmentioning

confidence: 99%

“…We also have discovered a new class of DNA adducts, 3-HA-dU, which are produced by the mutagenic and carcinogenic epoxides ethylene oxide (EO), propylene oxide (P0), glycidol, epichlorohydrin (ECH), and the epoxide of acrylonitrile (34)(35)(36)(37)(38). 3-HA-dU is formed after initial alkylation at the N3 position of dC, followed by rapid hydrolytic deamination to 3-HA-dU (34,37,38).…”

Section: Introductionmentioning

confidence: 99%

The Role of Mutagenic Metal Ions in Mediating In Vitro Mispairing by Alkylpyrimidines

Bhanot¹,

Solomon²

1994

Environmental Health Perspectives

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A variety of alkylating mutagens and carcinogens produce pyrimidine adducts in DNA that block DNA synthesis in vitro. Since DNA synthesis past the lesion is a necessary step to produce mutations, we investigated the role of the mutagenic metal ion Mn++ in facilitating DNA synthesis past alkylpyrimidines. In the presence of the natural metal activator Mg++, N3-ethyldeoxythymidine (N3-Et-dT) and 02-ethyideoxythymidine (02-Et-dT), present at a single site in DNA, blocked in vitro DNA synthesis 3' to the lesion and after incorporating dA opposite each lesion. The presence of Mn++ permitted postlesion synthesis with dT misincorporated opposite N3-Et-dT and 02-Et-dT, implicating these lesions in A.T->T-A transversion mutagenesis. The DNA synthesis block by in the presence of Mg++ was partial and was also removed by Mn++. Consistent with in vivo studies, dG was incorporated opposite 04-Et-dT during postlesion synthesis, leading to A-T-*G-C transition mutagenesis. We also have discovered a new class of DNA adducts, N3-hydroxyalkyldeoxyuridine (3-HA-dU) lesions, which are produced by mutagenic and carcinogenic aliphatic epoxides. 3-HA-dU is formed after initial alkylation at the N3 position of dC followed by a rapid hydrolytic deamination. As observed with the analogous mutagenic N3-Et-dT, the ethylene oxide-induced 3-hydroxyethyldeoxyuridine (3-HE-dU) blocked in vitro DNA synthesis, which could be bypassed in the presence of Mn++. The nucleotide incorporated opposite 3-HE-dU during postlesion synthesis is being identified. These studies suggest a role for Mn++ in mediating mutagenic and carcinogenic effects of environmentally important ethylating agents and aliphatic epoxides. -Environ Health Perspect 102(Suppl 3): 81-90 (1994).

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In vitro reaction of ethylene oxide with DNA and characterization of DNA adducts

Cited by 77 publications

References 44 publications

Inhalation Exposure of Rats and Mice to 1,3-Butadiene Induces N 6 -Adenine Adducts of Epoxybutene Detected by 32 P-Postlabeling and HPLC

Inhalation Exposure of Rats and Mice to 1,3-Butadiene Induces N 6 -Adenine Adducts of Epoxybutene Detected by 32 P-Postlabeling and HPLC

Dose-Response Relationships for N7-(2-Hydroxyethyl)Guanine Induced by Low-Dose [14C]Ethylene Oxide: Evidence for a Novel Mechanism of Endogenous Adduct Formation

The Role of Mutagenic Metal Ions in Mediating In Vitro Mispairing by Alkylpyrimidines

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