DNA polymerase action blocked by adenine adducts induced by 5-hydroxymethylchrysene sulfate.
Modification of M13mp10 single-stranded DNA with 5-hydroxymethylchrysene (5HCR) sulfate, the ultimate carcinogenic metabolite of 5-methylchrysene, resulted in formation of N6[(chrysen-5-yl)methyl]adenine and N2[(chrysen-5-yl)methyl]-guanine at the ratio of 2.7:1. Measurement of DNA synthesis using this modified template and E.coli DNA polymerase I (Klenow fragment) demonstrated that increasing levels of adducts caused a progressive decline in replication. Analysis of reaction products on DNA-sequence gels revealed DNA elongation to be arrested exclusively at adenine adducts in -AAAGGA- and -AACA- sequences.
Suzuki
M
M
Faculty of Pharmaceutical Science, Nagoya City University, Japan.
Kojima
M
M
Okuda
H
H
Watabe
T
T
Tada
M
M
D000225:Adenine / Q000378:metabolism; D001483:Base Sequence; D002273:Carcinogens / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D002911:Chrysenes / Q000378:metabolism*; D004247:DNA / Q000096:biosynthesis* / Q000378:metabolism; D004249:DNA Damage; D004256:DNA Polymerase I / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology; D006147:Guanine / Q000378:metabolism; D008969:Molecular Sequence Data
D002273:Carcinogens; D002911:Chrysenes; D006147:Guanine; C032670:5-hydroxymethylchrysene; D004247:DNA; D004256:DNA Polymerase I; D000225:Adenine