Alpha 2.0
Login Register
» Articles » PMID: 8319641

A Highly Sensitive and Specific Method for Quantitation of O-alkylated DNA Adducts and Its Application to the Analysis of Human Tissue DNA

Overview
Journal Environ Health Perspect
Date 1993 Mar 1
PMID 8319641
Citations 4
Authors
H Kang
C Konishi
T Kuroki
N Huh
Affiliations
Soon will be listed here.
Abstract

Formation and accumulation of O6-alkylguanine and O4-alkylthymine in human tissues is possibly the most relevant marker for cancer risk. Because humans are chronically exposed to diverse kinds of chemicals and eventual DNA structural modifications are supposed to be a complex mixture of adducts at very low levels, it is essential to use an assay with extremely high sensitivity and specificity. We have established a quantitation method, called PREPI, for O6-methylguanine, O4-methylthymine, and O4-ethylthymine by the combination of prefractionation by HPLC, 32P-postlabeling, and immunoprecipitation. The detection limit was about 1 fmole for all three adducts, enabling us to analyze about 1 x 10(-8) levels as a molar ratio to normal counterpart using 100 micrograms of DNA. In a pilot experiment, we analyzed 11 peripheral blood samples from healthy volunteers. O6-Methylguanine was detected in all the cases with a mean value of 2.0 +/- 1.3 x 10(-8) (range, 0.78-4.6 x 10(-8)). Neither O4-methylthymine nor O4-ethylthymine was above the detection limit of 0.8 x 10(-8) as a ratio to thymine.

Citing Articles

Proteins Associated with Neurodegenerative Diseases: Link to DNA Repair.

Khodyreva S, Dyrkheeva N, Lavrik O Biomedicines. 2025; 12(12.

PMID: 39767715 PMC: 11673744. DOI: 10.3390/biomedicines12122808.

In-Gene Quantification of O(6)-Methylguanine with Elongated Nucleoside Analogues on Gold Nanoprobes.

Trantakis I, Nilforoushan A, Dahlmann H, Stauble C, Sturla S J Am Chem Soc. 2016; 138(27):8497-504.

PMID: 27314828 PMC: 5726487. DOI: 10.1021/jacs.6b03599.

DNA repair mechanisms in dividing and non-dividing cells.

Iyama T, Wilson 3rd D DNA Repair (Amst). 2013; 12(8):620-36.

PMID: 23684800 PMC: 3720834. DOI: 10.1016/j.dnarep.2013.04.015.

Sensitivity of human type II topoisomerases to DNA damage: stimulation of enzyme-mediated DNA cleavage by abasic, oxidized and alkylated lesions.

Sabourin M, Osheroff N Nucleic Acids Res. 2000; 28(9):1947-54.

PMID: 10756196 PMC: 103304. DOI: 10.1093/nar/28.9.1947.

Influence of S-adenosylmethionine pool size on spontaneous mutation, dam methylation, and cell growth of Escherichia coli.

Posnick L, Samson L J Bacteriol. 1999; 181(21):6756-62.

PMID: 10542178 PMC: 94141. DOI: 10.1128/JB.181.21.6756-6762.1999.

References
1.
Umbenhauer D, Wild C, Montesano R, Saffhill R, Boyle J, Huh N . O(6)-methyldeoxyguanosine in oesophageal DNA among individuals at high risk of oesophageal cancer. Int J Cancer. 1985; 36(6):661-5. DOI: 10.1002/ijc.2910360607. View
2.
Huh N, Satoh M, Shiga J, Rajewsky M, Kuroki T . Immunoanalytical detection of O4-ethylthymine in liver DNA of individuals with or without malignant tumors. Cancer Res. 1989; 49(1):93-7. View
3.
Hall C, Badawi A, OConnor P, Saffhill R . The detection of alkylation damage in the DNA of human gastrointestinal tissues. Br J Cancer. 1991; 64(1):59-63. PMC: 1977320. DOI: 10.1038/bjc.1991.239. View
4.
Swenberg J, Dyroff M, Bedell M, Popp J, Huh N, Kirstein U . O4-ethyldeoxythymidine, but not O6-ethyldeoxyguanosine, accumulates in hepatocyte DNA of rats exposed continuously to diethylnitrosamine. Proc Natl Acad Sci U S A. 1984; 81(6):1692-5. PMC: 344984. DOI: 10.1073/pnas.81.6.1692. View
5.
Cameron V, Uhlenbeck O . 3'-Phosphatase activity in T4 polynucleotide kinase. Biochemistry. 1977; 16(23):5120-6. DOI: 10.1021/bi00642a027. View