Binding of benzo[a]pyrene at the 1,3,6 positions to nucleic acids in vivo on mouse skin and in vitro with rat liver microsomes and nuclei

Eleanor G Rogan, R. Roth, P. Katomski, J. Benderson, Ercole Cavalieri

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Loss of tritium from specific positions in [3H,14C] aromatic hydrocarbons can elucidate their binding site(s) to DNA and RNA and indicate the mechanism of activation. Studies of tritium loss from [6-3H,14C]benzo[a]pyrene(B[a]P), [1,3-3H,14C]B[a]P, [1,3,6-3H,14C]B[a]P, [6,7-3H,14C]B[a]P, and [7-3H,14C]B[a]P were conducted in vitro using liver nuclei and microsomes from 3-methylcholanthrene-induced Sprague-Dawley rats and in vivo on the skin of Charles River CD-1 mice. The relative loss of tritium from [3H, 14C]B[a]P was measured after binding to skin DNA and RNA, to nuclear DNA, and to native and denatured calf thymus and rat liver DNA's and poly(G) by microsomal activation. In skin, nuclei, and microsomes plus native DNA, virtually all B[a]P binding occurred at positions 1,3 and 6; while with microsomes plus denatured DNA or poly(G), B[a]P showed no binding at the 6 position and a small amount at the 1 and 3 positions. In vivo and with nuclei, binding at the 6 position predominated. Little loss of tritium from the 7 position was seen; this was expected because binding at this position is not thought to occur. This confirms the interpretation of loss of tritium as an indication of binding at a given position. These results demonstrate that the use of microsomes to activate B[a]P is not a valid model system for delineating the in vivo mechanism of B[a]P activation, and support previous evidence for one-electron oxidation as the mechanism of activation of hydrocarbons in binding to nucleic acids.

Original languageEnglish (US)
Pages (from-to)35-51
Number of pages17
JournalChemico-Biological Interactions
Volume22
Issue number1
DOIs
StatePublished - Jan 1 1978

Fingerprint

Benzo(a)pyrene
Liver Microsomes
Liver
Nucleic Acids
Rats
Skin
Tritium
DNA
Microsomes
Chemical activation
Poly G
In Vitro Techniques
RNA
Nuclear RNA
Thymus
Aromatic Hydrocarbons
Methylcholanthrene
Hydrocarbons
Rivers
Thymus Gland

ASJC Scopus subject areas

  • Toxicology

Cite this

Binding of benzo[a]pyrene at the 1,3,6 positions to nucleic acids in vivo on mouse skin and in vitro with rat liver microsomes and nuclei. / Rogan, Eleanor G; Roth, R.; Katomski, P.; Benderson, J.; Cavalieri, Ercole.

In: Chemico-Biological Interactions, Vol. 22, No. 1, 01.01.1978, p. 35-51.

Research output: Contribution to journalArticle

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abstract = "Loss of tritium from specific positions in [3H,14C] aromatic hydrocarbons can elucidate their binding site(s) to DNA and RNA and indicate the mechanism of activation. Studies of tritium loss from [6-3H,14C]benzo[a]pyrene(B[a]P), [1,3-3H,14C]B[a]P, [1,3,6-3H,14C]B[a]P, [6,7-3H,14C]B[a]P, and [7-3H,14C]B[a]P were conducted in vitro using liver nuclei and microsomes from 3-methylcholanthrene-induced Sprague-Dawley rats and in vivo on the skin of Charles River CD-1 mice. The relative loss of tritium from [3H, 14C]B[a]P was measured after binding to skin DNA and RNA, to nuclear DNA, and to native and denatured calf thymus and rat liver DNA's and poly(G) by microsomal activation. In skin, nuclei, and microsomes plus native DNA, virtually all B[a]P binding occurred at positions 1,3 and 6; while with microsomes plus denatured DNA or poly(G), B[a]P showed no binding at the 6 position and a small amount at the 1 and 3 positions. In vivo and with nuclei, binding at the 6 position predominated. Little loss of tritium from the 7 position was seen; this was expected because binding at this position is not thought to occur. This confirms the interpretation of loss of tritium as an indication of binding at a given position. These results demonstrate that the use of microsomes to activate B[a]P is not a valid model system for delineating the in vivo mechanism of B[a]P activation, and support previous evidence for one-electron oxidation as the mechanism of activation of hydrocarbons in binding to nucleic acids.",
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