DNA polymerase action blocked by adenine adducts induced by 5-hydroxymethylchrysene sulfate

Biochem Int. 1991 Sep;25(1):19-27.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine / metabolism
  • Base Sequence
  • Carcinogens / metabolism*
  • Chromatography, High Pressure Liquid
  • Chrysenes / metabolism*
  • DNA / biosynthesis*
  • DNA / metabolism
  • DNA Damage*
  • DNA Polymerase I / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / enzymology
  • Guanine / metabolism
  • Molecular Sequence Data

Substances

  • Carcinogens
  • Chrysenes
  • Guanine
  • 5-hydroxymethylchrysene
  • DNA
  • DNA Polymerase I
  • Adenine