Mutagenic spectrum of butadiene-derived N1-deoxyinosine adducts and N6,N6-deoxyadenosine intrastrand cross-links in mammalian cells

Manorama Kanuri, Lubomir V. Nechev, Pamela J. Tamura, Constance M. Harris, Thomas M. Harris, Robert (Stephen) Lloyd

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Reactive metabolites of 1,3-butadiene, including 1,2-epoxy-3-butene (BDO), 1,2:3,4-diepoxybutane (BDO2), and 3,4-epoxy-1,2-butanediol (BDE), form both stable and unstable base adducts in DNA and have been implicated in producing genotoxic effects in rodents and human cells. N1 deoxyadenosine adducts are unstable and can undergo either hydrolytic deamination to yield N1 deoxyinosine adducts or Dimroth rearrangement to yield N6 adducts. The dominant point mutation observed at AT sites in both in vivo and in vitro mutagenesis studies using BD and its epoxides has been A → T transversions followed by A → G transitions. To understand which of the butadiene adducts are responsible for mutations at AT sites, the present study focuses on the N1 deoxyinosine adduct at C2 of BDO and N6,N6-deoxyadenosine intrastrand cross-links derived from BDO2. These lesions were incorporated site-specifically and stereospecifically into oligodeoxynucleotides which were engineered into mammalian shuttle vectors for replication bypass and mutational analyses in COS-7 cells. Replication of DNAs containing the R,R-BDO2 intrastrand cross-link between N6 positions of deoxyadenosine yielded a high frequency (59%) of single base substitutions at the 3′ adducted base, while 19% mutagenesis was detected using the S,S-diastereomer. Comparable studies using the R- and S-diastereomers of the N1 deoxyinosine adduct gave rise to ∼50 and 80% A → G transitions with overall mutagenic frequencies of 59 and 90%, respectively. Collectively, these data establish a molecular basis for A → G transitions that are observed following in vivo and in vitro exposures to BD and its epoxides, but fail to reveal the source of the A → T transversions that are the dominant point mutation.

Original languageEnglish (US)
Pages (from-to)1572-1580
Number of pages9
JournalChemical Research in Toxicology
Volume15
Issue number12
DOIs
StatePublished - Dec 1 2002
Externally publishedYes

Fingerprint

Mutagenesis
Epoxy Compounds
Cells
Point Mutation
Genetic Vectors
Deamination
DNA Adducts
Oligodeoxyribonucleotides
COS Cells
DNA
Metabolites
DNA Replication
Rodentia
Substitution reactions
Mutation
deoxyinosine
erythritol anhydride
1,3-butadiene
2'-deoxyadenosine
In Vitro Techniques

ASJC Scopus subject areas

  • Drug Discovery
  • Organic Chemistry
  • Chemistry(all)
  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Mutagenic spectrum of butadiene-derived N1-deoxyinosine adducts and N6,N6-deoxyadenosine intrastrand cross-links in mammalian cells. / Kanuri, Manorama; Nechev, Lubomir V.; Tamura, Pamela J.; Harris, Constance M.; Harris, Thomas M.; Lloyd, Robert (Stephen).

In: Chemical Research in Toxicology, Vol. 15, No. 12, 01.12.2002, p. 1572-1580.

Research output: Contribution to journalArticle

Kanuri, Manorama ; Nechev, Lubomir V. ; Tamura, Pamela J. ; Harris, Constance M. ; Harris, Thomas M. ; Lloyd, Robert (Stephen). / Mutagenic spectrum of butadiene-derived N1-deoxyinosine adducts and N6,N6-deoxyadenosine intrastrand cross-links in mammalian cells. In: Chemical Research in Toxicology. 2002 ; Vol. 15, No. 12. pp. 1572-1580.
@article{e49f7e0fb6ba4b1d99ee74399484d310,
title = "Mutagenic spectrum of butadiene-derived N1-deoxyinosine adducts and N6,N6-deoxyadenosine intrastrand cross-links in mammalian cells",
abstract = "Reactive metabolites of 1,3-butadiene, including 1,2-epoxy-3-butene (BDO), 1,2:3,4-diepoxybutane (BDO2), and 3,4-epoxy-1,2-butanediol (BDE), form both stable and unstable base adducts in DNA and have been implicated in producing genotoxic effects in rodents and human cells. N1 deoxyadenosine adducts are unstable and can undergo either hydrolytic deamination to yield N1 deoxyinosine adducts or Dimroth rearrangement to yield N6 adducts. The dominant point mutation observed at AT sites in both in vivo and in vitro mutagenesis studies using BD and its epoxides has been A → T transversions followed by A → G transitions. To understand which of the butadiene adducts are responsible for mutations at AT sites, the present study focuses on the N1 deoxyinosine adduct at C2 of BDO and N6,N6-deoxyadenosine intrastrand cross-links derived from BDO2. These lesions were incorporated site-specifically and stereospecifically into oligodeoxynucleotides which were engineered into mammalian shuttle vectors for replication bypass and mutational analyses in COS-7 cells. Replication of DNAs containing the R,R-BDO2 intrastrand cross-link between N6 positions of deoxyadenosine yielded a high frequency (59{\%}) of single base substitutions at the 3′ adducted base, while 19{\%} mutagenesis was detected using the S,S-diastereomer. Comparable studies using the R- and S-diastereomers of the N1 deoxyinosine adduct gave rise to ∼50 and 80{\%} A → G transitions with overall mutagenic frequencies of 59 and 90{\%}, respectively. Collectively, these data establish a molecular basis for A → G transitions that are observed following in vivo and in vitro exposures to BD and its epoxides, but fail to reveal the source of the A → T transversions that are the dominant point mutation.",
author = "Manorama Kanuri and Nechev, {Lubomir V.} and Tamura, {Pamela J.} and Harris, {Constance M.} and Harris, {Thomas M.} and Lloyd, {Robert (Stephen)}",
year = "2002",
month = "12",
day = "1",
doi = "10.1021/tx025591g",
language = "English (US)",
volume = "15",
pages = "1572--1580",
journal = "Chemical Research in Toxicology",
issn = "0893-228X",
publisher = "American Chemical Society",
number = "12",

}

TY - JOUR

T1 - Mutagenic spectrum of butadiene-derived N1-deoxyinosine adducts and N6,N6-deoxyadenosine intrastrand cross-links in mammalian cells

AU - Kanuri, Manorama

AU - Nechev, Lubomir V.

AU - Tamura, Pamela J.

AU - Harris, Constance M.

AU - Harris, Thomas M.

AU - Lloyd, Robert (Stephen)

PY - 2002/12/1

Y1 - 2002/12/1

N2 - Reactive metabolites of 1,3-butadiene, including 1,2-epoxy-3-butene (BDO), 1,2:3,4-diepoxybutane (BDO2), and 3,4-epoxy-1,2-butanediol (BDE), form both stable and unstable base adducts in DNA and have been implicated in producing genotoxic effects in rodents and human cells. N1 deoxyadenosine adducts are unstable and can undergo either hydrolytic deamination to yield N1 deoxyinosine adducts or Dimroth rearrangement to yield N6 adducts. The dominant point mutation observed at AT sites in both in vivo and in vitro mutagenesis studies using BD and its epoxides has been A → T transversions followed by A → G transitions. To understand which of the butadiene adducts are responsible for mutations at AT sites, the present study focuses on the N1 deoxyinosine adduct at C2 of BDO and N6,N6-deoxyadenosine intrastrand cross-links derived from BDO2. These lesions were incorporated site-specifically and stereospecifically into oligodeoxynucleotides which were engineered into mammalian shuttle vectors for replication bypass and mutational analyses in COS-7 cells. Replication of DNAs containing the R,R-BDO2 intrastrand cross-link between N6 positions of deoxyadenosine yielded a high frequency (59%) of single base substitutions at the 3′ adducted base, while 19% mutagenesis was detected using the S,S-diastereomer. Comparable studies using the R- and S-diastereomers of the N1 deoxyinosine adduct gave rise to ∼50 and 80% A → G transitions with overall mutagenic frequencies of 59 and 90%, respectively. Collectively, these data establish a molecular basis for A → G transitions that are observed following in vivo and in vitro exposures to BD and its epoxides, but fail to reveal the source of the A → T transversions that are the dominant point mutation.

AB - Reactive metabolites of 1,3-butadiene, including 1,2-epoxy-3-butene (BDO), 1,2:3,4-diepoxybutane (BDO2), and 3,4-epoxy-1,2-butanediol (BDE), form both stable and unstable base adducts in DNA and have been implicated in producing genotoxic effects in rodents and human cells. N1 deoxyadenosine adducts are unstable and can undergo either hydrolytic deamination to yield N1 deoxyinosine adducts or Dimroth rearrangement to yield N6 adducts. The dominant point mutation observed at AT sites in both in vivo and in vitro mutagenesis studies using BD and its epoxides has been A → T transversions followed by A → G transitions. To understand which of the butadiene adducts are responsible for mutations at AT sites, the present study focuses on the N1 deoxyinosine adduct at C2 of BDO and N6,N6-deoxyadenosine intrastrand cross-links derived from BDO2. These lesions were incorporated site-specifically and stereospecifically into oligodeoxynucleotides which were engineered into mammalian shuttle vectors for replication bypass and mutational analyses in COS-7 cells. Replication of DNAs containing the R,R-BDO2 intrastrand cross-link between N6 positions of deoxyadenosine yielded a high frequency (59%) of single base substitutions at the 3′ adducted base, while 19% mutagenesis was detected using the S,S-diastereomer. Comparable studies using the R- and S-diastereomers of the N1 deoxyinosine adduct gave rise to ∼50 and 80% A → G transitions with overall mutagenic frequencies of 59 and 90%, respectively. Collectively, these data establish a molecular basis for A → G transitions that are observed following in vivo and in vitro exposures to BD and its epoxides, but fail to reveal the source of the A → T transversions that are the dominant point mutation.

UR - http://www.scopus.com/inward/record.url?scp=12244301638&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=12244301638&partnerID=8YFLogxK

U2 - 10.1021/tx025591g

DO - 10.1021/tx025591g

M3 - Article

VL - 15

SP - 1572

EP - 1580

JO - Chemical Research in Toxicology

JF - Chemical Research in Toxicology

SN - 0893-228X

IS - 12

ER -