/sup 32/P-postlabeling analysis of non-radioactive aromatic carcinogen-DNA adducts

Journal Article · Fri Jan 01 04:00:00 EST 1982 · Carcinogenesis (N.Y.); (United States)

DOI:https://doi.org/10.1093/carcin/3.9.1081· OSTI ID:6120642

Gupta, R C; Reddy, M V; Randerath, K ^[1]

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1. Texas Medical Center, Houston (USA)

A newly developed enzymatic /sup 32/P-postlabeling method was applied to the analysis of DNA's containing non-radioactive arylamine, arylamide, and polycyclic aromatic hydrocarbon adducts. DNA reacted in vitro with N-hydroxy-2-amino-fluorene, N-acetoxy-2-acetylaminofluorene, and 7..beta..,8..cap alpha..-dihydroxy-9..cap alpha.., 10..cap alpha..-epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene, respectively, as well as DNA preparations from the livers of rats treated with N-hydroxy-2-acetylaminofluorene and benzo(a)pyrene, respectively, were enzymatically digested to deoxyribonucleoside 3'-monophosphates, which were then converted to (5'-/sup 32/P)deoxyribonucleoside 3',5'-bisphosphates by T4 polynucleotide kinase-catalyzed (/sup 32/P)phosphate transfer from (..gamma..-/sup 32/P)ATP. The /sup 32/P-labeled nucleotides were resolved by anion-exchange t.l.c. on polyethyleneimine-cellulose and detected by autoradiography. Aromatic adduct nucleotides were found to be retained at the origin in aqueous electrolyte solutions but to migrate as distinct spots in solvents containing 7-8.5 M urea. Advantage was taken of this observation to remove /sup 32/P-labeled normal DNA nucleotides from adduct nucleotides. This purification enabled the detection of a single adduct in 10/sup 7/-10/sup 8/ normal nucleotides. The method appears applicable to the ultrasensitive detection of a large number of carcinogen-DNA adducts of diverse structure without requiring radioactive carcinogens or specific antibodies.

• OSTI ID: 6120642
• Journal Information: Carcinogenesis (N.Y.); (United States), Journal Name: Carcinogenesis (N.Y.); (United States) Vol. 3:9; ISSN CRNGD
• Country of Publication: United States
• Language: English

Related Subjects

550901* -- Pathology-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADDUCTS
ANIMALS
AROMATICS
BENZOPYRENE
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CALVES
CARCINOGENS
CATTLE
CONDENSED AROMATICS
DAYS LIVING RADIOISOTOPES
DETECTION
DIGESTIVE SYSTEM
DNA
DOMESTIC ANIMALS
GLANDS
HYDROCARBONS
IN VITRO
IN VIVO
ISOTOPE APPLICATIONS
ISOTOPES
LABELLING
LIGHT NUCLEI
LIVER
LYMPHATIC SYSTEM
MAMMALS
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
RADIOENZYMATIC ASSAY
RADIOISOTOPES
RATS
RODENTS
RUMINANTS
THYMUS
TRACER TECHNIQUES
VERTEBRATES