Covalent Binding of Benzo[a]pyrene to Free and Membrane-Bound DNA in Isolated Liver Nuclei from 3-Methylcholanthrene-Induced Rats

Eleanor G Rogan, Sheila Higginbotham, Prabhakar D. Devanesan, Ercole Cavalieri

Research output: Contribution to journalArticle

Abstract

Benzo[a]pyrene (BP) is bound to DNA by two major pathways: one-electron oxidation and monooxygenation. One-electron oxidation requires that the highly reactive radical cation be formed in close proximity to the DNA. Thus, in intact cell nuclei, binding of BP radical cation would be expected to occur predominantly in DNA associated with the membrane. To examine this, nuclei from the livers of 3-methylcholanthrene-induced male MRC Wistar rats were incubated with [3H]BP and NADPH. After the incubation, the “free” DNA was separated from “membrane-bound” DNA by extraction in low-Mg buffer. Binding of BP to membrane-bound DNA was 9.5 ± 2.0 µmol/mol DNA-P, whereas binding to free DNA was 2.2 ± 0.7 µmol/mol DNA-P. BP adducts formed by one-electron oxidation and lost from the nuclear DNA by depurination were also examined. The three depurination adducts previously found with rat liver microsomes and in mouse skin were also obtained from nuclei. These results suggest that binding of BP to DNA in intact nuclei occurs preferentially to DNA associated with the nuclear membrane.

Original languageEnglish (US)
Pages (from-to)119-126
Number of pages8
JournalPolycyclic Aromatic Compounds
Volume6
Issue number1-4
DOIs
StatePublished - Jan 1994

Fingerprint

Methylcholanthrene
Benzo(a)pyrene
Pyrene
Liver
Rats
DNA
Membranes
Oxidation
Cations
Electrons
Positive ions
NADP
Skin
Buffers
Cells

Keywords

  • DNA adducts
  • benzo[a]pyrene
  • one-electron oxidation
  • radical cations
  • rat liver nuclei

ASJC Scopus subject areas

  • Polymers and Plastics
  • Organic Chemistry
  • Materials Chemistry

Cite this

Covalent Binding of Benzo[a]pyrene to Free and Membrane-Bound DNA in Isolated Liver Nuclei from 3-Methylcholanthrene-Induced Rats. / Rogan, Eleanor G; Higginbotham, Sheila; Devanesan, Prabhakar D.; Cavalieri, Ercole.

In: Polycyclic Aromatic Compounds, Vol. 6, No. 1-4, 01.1994, p. 119-126.

Research output: Contribution to journalArticle

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