32P-postlabeling detection of DNA adducts in fish from chemically contaminated waterways.
Fish were collected from sites in the chemically-contaminated Buffalo River, New York, and the Detroit River, Michigan. The sediments of these rivers have high levels of chemical contaminants, including polycyclic aromatic hydrocarbons (PAHs), and fish from these locations have high prevalences of liver cancer. To determine chemical-DNA interactions and a possible role for chemicals as a cause of the observed tumors, DNA was isolated from livers and was enzymatically digested to normal and adducted nucleotides. The DNA digests were enriched for hydrophobic, bulky adducts, either by preparative reverse phase high pressure liquid chromatography, or by selective nuclease P1 dephosphorylation of normal nucleotides. DNA-chemical adducts were then quantitated by 32P-postlabeling analysis. Regardless of the adduct enrichment procedure, the chromatograms derived from DNA of fish from polluted areas showed a diffuse, diagonal radioactive zone consisting, at least in part, of multiple overlapping discrete adduct spots. The behavior of the adducts in the diagonal radioactive zone and of their unlabeled precursors is consistent with their identification as nucleotide adducts of a variety of bulky, hydrophobic, aromatic genotoxic compounds. Analysis of bile demonstrated recent exposure to multi-ringed aromatic compounds.
Roswell Park Memorial Institute, Buffalo, New York 142163.
A E
AE
Maccubbin
J J
JJ
Black
B P
BP
Dunn
D000818:Animals; D001345:Autoradiography; D001646:Bile / Q000378:metabolism; D002347:Carps / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D005399:Fishes / Q000378:metabolism*; D005618:Fresh Water; D008099:Liver / Q000378:metabolism / Q000473:pathology*; D008824:Michigan; D009518:New York; D010761:Phosphorus Radioisotopes; D011083:Polycyclic Compounds / Q000378:metabolism* / Q000633:toxicity; D011865:Radioisotope Dilution Technique; D013045:Species Specificity; D014873:Water Pollutants / Q000633:toxicity; D014874:Water Pollutants, Chemical / Q000633:toxicity
D010761:Phosphorus Radioisotopes; D011083:Polycyclic Compounds; D014873:Water Pollutants; D014874:Water Pollutants, Chemical; D004247:DNA